CN104231023B

CN104231023B - Tricyclic fused heterocycle nucleoside phosphoramidate compound, preparation method and application

Info

Publication number: CN104231023B
Application number: CN201410245982.0A
Authority: CN
Inventors: 王勇; 赵立文; 毕胜; 高毅平; 王德忠; 陈宏雁; 刘阳; 张仓; 张文萍; 南阳
Original assignee: Nanjing Huicheng Pharmaceutical Co Ltd
Current assignee: Nanjing Huicheng Pharmaceutical Co., Ltd.
Priority date: 2013-06-06
Filing date: 2014-06-04
Publication date: 2019-02-05
Anticipated expiration: 2034-06-04
Also published as: WO2014194826A1; CN104231023A

Abstract

The present invention provides a kind of tricyclic fused heterocycle nucleoside phosphoramidate compounds, preparation method, the composition containing the nucleoside phosphoramidate compound, and purposes of the compound or composition as disease of viral infection therapeutic agent, especially as the purposes of viral hepatitis treatment drug.

Description

Tricyclic fused heterocycle nucleoside phosphoramidate compound, preparation method and application

Technical field

The invention belongs to medicinal chemistry arts, and in particular to a kind of tricyclic fused heterocycle nucleoside phosphoramidate compound, Preparation method, the composition containing the nucleoside phosphoramidate compound and the compound or composition are as virus The purposes of infectious diseases therapeutic agent, especially as the purposes of viral hepatitis treatment drug.

Background technique

Hepatitis C Virus (HCV) is the main pathogens for causing most of non-A non-B hepatitis.Hepatitis C Virus Infection is to lead to chronic liver disease, such as the major health concern of cirrhosis and liver cancer.According to the data of the World Health Organization, the whole world 200,000,000 infected population is had more than, once infection, only about 20% people can remove virus, others will carry HCV.

HCV is a kind of virus for belonging to flaviviridae, is to have enveloped positive strand RNA virus in flaviviridae family.Single-stranded HCV About 9500 nucleotide of the length of rna gene group, (ORF) containing single open reading frame encode the big poly- of about 3000 amino acid Hop protein.

It is that the gene structure of pestivirus and Flavivirus is similar that HCV belongs to other two in flaviviridae.Currently, controlling The standard method for treating HCV infection has interferon and interferon and ribavirin combination therapy.But, only 50% curer couple This method has a reaction, and interferon has an apparent side effect, such as flu-like symptoms, weight lower and tired It is powerless, and interferon and ribavirin combination therapy then generate sizable side effect, including haemolysis, anemia and tired.

For a long time, due to the cell model of the suitable external virus replication of shortage, novel anti-HCV medicament progress is developed Slowly.Until 1999, Lohmann established the sub-genome duplication model that high-level can independently replicate in liver cancer cells (replicon), the model are selective bicistronic mRNA subgenome HCV RNA replicon system, are acknowledged as HCV RNA The research of replication in vitro model obtains the milestone of breakthrough development.Replicon can high level in human hepatoma cell strain Huh7 cell strain Autonomous duplication.Research finds that replicon replicates in cell and has no significant effect to host cell growth and metabolism that high level is multiple It makes related with the receptivity of virus genomic adaptive mutation and host cell.These results of study are selectivity HCV total order The research of column subgenomic replicons is laid a good foundation.

Summary of the invention

It is an object of the present invention to provide thick for treating and/or preventing tricyclic shown in the logical formula (I) of HCV infection Heterocyclic nucleoside phosphoramidate compound, its stereoisomer, pro-drug, pharmaceutically acceptable salt, hydrate, solvent Object is closed,

P* indicates chiral phosphorus atoms,

Wherein, (1) R₁Selected from H and alkyl, the alkyl is optionally by one or more alkyl, alkoxy, alkylamino, halogen Element, hydroxyl, amino, nitro, cyano, alkyl acyl, aminoacyl, alkylamino acyl group, sulfonyl, sulfinyl, sulfydryl, aryl Or heteroaryl replaces；

(2)R₂Selected from H, alkyl, aminoacyl, aryl and heteroaryl, the alkyl is optionally by one or more alkyl, alkane Oxygroup, halogen, hydroxyl, amino, single alkylamino, double alkylaminos, aryl or heteroaryl replace；

(3)R₃For there is no, or be selected from alkyl, alkenyl, alkynyl, alkoxy, halogen, halogenated alkyl, alkyl amino, alkyl Sulfonyl, alkyl sulfonyl amino, sulfamoyl alkyl, halogenated alkoxy, hydroxyl, nitro, amino, single alkylamino, double alkylaminos, Acylamino-, ester group ,-CN, naphthenic base, Heterocyclylalkyl, aryl, heteroaryl, aralkyl；

(4) D is selected from-(CR_a1R_a2)_mO-、-(CR_a1R_a2)_mS-、-SO-、-SO₂-、-CO-、-N(R_b)-、-N(R_b)-CO-、-C (R_c1R_c2)-, is wherein the R_a1、R_a2Respectively hydrogen, alkyl, halogen, halogenated alkyl, alkenyl or halogenated alkenyl or R_a1、R_a2 C in connection constitutes naphthenic base together, and the m is 0 or 1, the R_bTo be not present, hydrogen, alkyl, alkyl sulphonyl Or alkyl-carbonyl, the R_c1、R_c2Respectively hydrogen, halogen, alkyl, halogenated alkyl, alkenyl or halogenated alkenyl, or work as R_c1、R_c2? When for alkyl, C can be with R_c1、R_c2Constitute naphthenic base；

(5)D¹Selected from oxygen, sulphur ,-SO- ,-SO₂-、-CO-、-N(R_d)-、-(CH)_n, wherein the R_dTo be not present, hydrogen, Alkyl or halogenated alkyl, the n are 0,1 or 2, described-(CH)_nOptionally by alkoxy, alkylamino, halogen, hydroxyl, ammonia Base, nitro, cyano, alkyl acyl, aminoacyl, alkylamino acyl group, sulfonyl, sulfinyl, sulfydryl, aryl or heteroaryl take Generation；

(6)G、G¹Respectively-N- or-CH-；

(7)For singly-bound or double bond, two of themIt is singly-bound or oneFor singly-bound, and it is another One is double bond；With

(8) ring T is selected from:

A, aromatic ring or hetero-aromatic ring, the aromatic ring or hetero-aromatic ring can be by R₄Replace, wherein R₄For alkyl, alkenyl, alkynyl, Alkoxy, halogen, halogenated alkyl, halogenated alkoxy, hydroxyl, nitro, amino, single alkylamino, double alkylaminos, alkyl acyl, ammonia Base acyl group, alkylamino acyl group, acylamino- ,-CN, aryl or heteroaryl；With

B, cycloalkane or heterocycloalkane, the cycloalkane or heterocycloalkane can be by R₄Replace, wherein R₄For alkyl, alkene Base, alkynyl, alkoxy, halogen, halogenated alkyl, halogenated alkoxy, hydroxyl, nitro, amino, single alkylamino, double alkylaminos, alkyl Acyl group, aminoacyl, alkylamino acyl group, acylamino- ,-CN, aryl or heteroaryl.It is another object of the present invention to provide systems It is the nucleoside phosphoramidate compound of standby logical formula (I) of the invention, its stereoisomer, pro-drug, pharmaceutically acceptable Salt, hydrate, solvate or the method for crystallization.

It is yet a further object of the present invention to provide the nucleoside phosphoramidate compound comprising logical formula (I) of the invention, The group of its stereoisomer, pro-drug, pharmaceutically acceptable salt, hydrate or solvate and the acceptable carrier of drug effect Close object and nucleoside phosphoramidate compound, its stereoisomer, pro-drug, pharmacy comprising logical formula (I) of the invention The composition of upper acceptable salt, hydrate or solvate and another antiviral agent.

Another purpose of the invention is that providing, the nucleoside phosphoramidate compound of logical formula (I) of the invention, it is vertical Body isomers, pro-drug, pharmaceutically acceptable salt, hydrate, solvate treatment and/or prevention of hepatitis C sense It is the nucleoside phosphoramidate compound and its stereoisomer of the method for dye and logical formula (I) of the invention, salt, hydrate, molten Agent close object or crystallization preparation for treat and/or the drug of prevention of hepatitis C infection in application.

The further object of the present invention, which is to provide, inhibits RNA Dependent RNA varial polymerases, especially inhibition HCVNS5B The method of polymerase.

The object of the invention is also to provide in treatment RNA Dependent RNA virus replication, especially treatment HCV duplication Application.

Another object of the present invention also resides in offer for treating RNA Dependent RNA virus infection, especially treats Application in HCV infection.

For foregoing invention purpose, the present invention the following technical schemes are provided:

In a first aspect, the present invention provide a kind of tricyclic fused heterocycle nucleoside phosphoramidate compound, its stereoisomer, Pro-drug, pharmaceutically acceptable salt, hydrate, solvate or crystallization, the compound is as shown in logical formula (I):

P* indicates chiral phosphorus atoms,

Wherein,

(1)R₁Selected from H and alkyl, the alkyl is optionally by one or more alkyl, alkoxy, alkylamino, halogen, hydroxyl Base, amino, nitro, cyano, alkyl acyl, aminoacyl, alkylamino acyl group, sulfonyl, sulfinyl, sulfydryl, aryl or miscellaneous Aryl replaces；

(6)G、G¹Respectively-N- or-CH-；

(7)For singly-bound or double bond, two of themIt is singly-bound or oneFor singly-bound, and Another is double bond；With

(8) ring T is selected from:

B, cycloalkane or heterocycloalkane, the cycloalkane or heterocycloalkane can be by R₄Replace, wherein R₄For alkyl, alkene Base, alkynyl, alkoxy, halogen, halogenated alkyl, halogenated alkoxy, hydroxyl, nitro, amino, single alkylamino, double alkylaminos, alkyl Acyl group, aminoacyl, alkylamino acyl group, acylamino- ,-CN, aryl or heteroaryl.

In some embodiments of first aspect, the present invention provides the nucleoside phosphoramidate chemical combination of above-mentioned logical formula (I) Object, its stereoisomer, pro-drug, pharmaceutically acceptable salt, hydrate, solvate or crystallization, wherein

D is selected from-(CR_a1R_a2)_mO-、-(CR_a1R_a2)_mS-、-SO-、-SO₂-、-CO-、-N(R_b)-、-N(R_b)-CO-、-C (R_c1R_c2)-, is wherein the R_a1、R_a2Respectively hydrogen, C_1-6Alkyl, halogen, halogenated C_1-6Alkyl, C_2-6Alkenyl or halogenated C_2-6Alkene Base or R_a1、R_a2C in connection constitutes C together_3-8Naphthenic base, the m are 0 or 1, the R_bTo be not present, hydrogen, C_1-6Alkyl, C_1-6Alkyl sulphonyl or C_1-6Alkyl-carbonyl, the R_c1、R_c2Respectively hydrogen, halogen, C_1-6Alkyl, halogenated C_1-6 Alkyl, C_2-6Alkenyl or halogenated C_2-6Alkenyl, or work as R_c1、R_c2When being alkyl, C can be with R_c1、R_c2Constitute C_3-8Naphthenic base；

D¹Selected from oxygen, sulphur ,-SO- ,-SO₂-、-CO-、-N(R_d)-、-(CH)_n, wherein the R_dTo be not present, hydrogen, C_1-6 Alkyl, halogenated C_1-6Alkyl, the n are 0,1 or 2, described-(CH)_nOptionally by C_1-6Alkoxy, C_1-6Alkylamino, halogen, Hydroxyl, amino, nitro, cyano, C_1-6Alkyl acyl, aminoacyl, C_1-6Alkylamino acyl group, sulfonyl, sulfinyl, sulfydryl, Aryl or heteroaryl replace；With

Ring T is selected from:

a、C_5-10Aromatic ring or hetero-aromatic ring, the aromatic ring or hetero-aromatic ring can be by R₄Replace, wherein R₄For C_1-6Alkyl, C_2-6 Alkenyl, C_2-6Alkynyl, C_1-6Alkoxy, halogen, halogenated C_1-6Alkyl, halogenated C_1-6Alkoxy, hydroxyl, nitro, amino, list C_1-6Alkane Amino, double C_1-6Alkylamino, C_1-6Alkyl acyl, aminoacyl, C_1-6Alkylamino acyl group, acylamino- ,-CN, aryl or heteroaryl； With

b、C_3-10Cycloalkane or heterocycloalkane, the cycloalkane or heterocycloalkane can be by R₄Replace, wherein R₄For C_1-6 Alkyl, C_2-6Alkenyl, C_2-6Alkynyl, C_1-6Alkoxy, halogen, halogenated C_1-6Alkyl, halogenated C_1-6Alkoxy, hydroxyl, nitro, amino, Single C_1-6Alkylamino, double C_1-6Alkylamino, C_1-6Alkyl acyl, aminoacyl, C_1-6Alkylamino acyl group, acylamino- ,-CN, aryl or Heteroaryl.

In other embodiments of first aspect, the present invention provides the nucleoside phosphoramidate of above-mentioned logical formula (I) Object, its stereoisomer, pro-drug, pharmaceutically acceptable salt, hydrate, solvate or crystallization are closed, wherein

R₁Selected from H and C_1-6Alkyl, the alkyl is optionally by one or more C_1-6Alkyl, C_1-6Alkoxy, C_1-6Alkylamino, Halogen, hydroxyl, amino, nitro, cyano, C_1-6Alkyl acyl, aminoacyl, C_1-6Alkylamino acyl group, sulfonyl, sulfinyl, Sulfydryl, aryl or heteroaryl replace；

R₂Selected from H, C_1-6Alkyl, aminoacyl, aryl and heteroaryl, the alkyl is optionally by one or more C_1-6Alkane Base, C_1-6Alkoxy, halogen, hydroxyl, amino, list C_1-6Alkylamino, double C_1-6Alkylamino, aryl or heteroaryl replace；With

R₃For there is no, or be selected from C_1-6Alkyl, C_2-6Alkenyl, C_2-6Alkynyl, C_1-6Alkoxy, halogen, halogenated C_1-6Alkyl, C_1-6Alkyl amino, C_1-6Alkyl sulphonyl, C_1-6Alkyl sulfonyl amino, sulfamoyl C_1-6Alkyl, halogenated C_1-6Alkoxy, hydroxyl, Nitro, amino, list C_1-6Alkylamino, double C_1-6Alkylamino, acylamino-, ester group ,-CN, C_3-8Naphthenic base, C_3-8Heterocyclylalkyl, aryl, Heteroaryl, virtue C_1-6Alkyl.

(1)R₁Selected from H and C_1-6Alkyl, the alkyl is optionally by one or more C_1-6Alkoxy, C_1-6Alkylamino, halogen, Hydroxyl, amino, nitro, cyano, C_1-6Alkyl acyl, aminoacyl, C_1-6Alkylamino acyl group, sulfonyl, sulfinyl, sulfydryl, Aryl or heteroaryl replace；

(2)R₂Selected from H, C_1-6Alkyl, aminoacyl, aryl and heteroaryl, the alkyl is optionally by one or more C_1-6 Alkoxy, halogen, hydroxyl, amino, list C_1-6Alkylamino, double C_1-6Alkylamino, aryl or heteroaryl replace；

(3)R₃For there is no, or be selected from C_1-6Alkyl, C_1-6Alkoxy, halogen, halogenated C_1-6Alkyl, halogenated C_1-6Alkoxy, Hydroxyl, nitro, amino, list C_1-6Alkylamino, double C_1-6Alkylamino, acylamino-, ester group ,-CN, C_3-8Naphthenic base, miscellaneous C_3-8Naphthenic base, Aryl, heteroaryl, virtue C_1-6Alkyl；

(4) D is selected from-(CR_a1R_a2)_mO-、-(CR_a1R_a2)_mS-、-SO-、-SO₂-、-CO-、-N(R_b)-、-N(R_b)-CO-、-C (R_c1R_c2)-, is wherein the R_a1、R_a2Respectively hydrogen, C_1-6Alkyl, halogen, halogenated C_1-6Alkyl, C_2-6Alkenyl or halogenated C_2-6Alkene Base or R_a1、R_a2C in connection constitutes C together_3-8Naphthenic base, the m are 0 or 1, the R_bTo be not present, hydrogen, Alkyl, alkyl sulphonyl or alkyl-carbonyl, the R_c1、R_c2Respectively hydrogen, halogen, alkyl, alkenyl or halogenated alkenyl, or work as R_c1、R_c2When being alkyl, C can be with R_c1、R_c2Constitute naphthenic base；

(5)D¹Selected from oxygen, sulphur ,-SO- ,-SO₂-、-CO-、-N(R_d)-、-(CH)_n, the R_dTo be not present, hydrogen, C_1-6 Alkyl or halogenated C_1-6Alkyl, the n are 0,1 or 2, described-(CH)_nOptionally by C_1-6Alkoxy, C_1-6Alkylamino, halogen Element, hydroxyl, amino, nitro, cyano, C_1-6Alkyl acyl, aminoacyl, C_1-6Alkylamino acyl group, sulfonyl, sulfinyl, mercapto Base, aryl or heteroaryl replace；

(6)G、G¹Respectively-N- or-CH-；

(7)For singly-bound or double bond, two of themIt is singly-bound or oneFor singly-bound, and it is another One is double bond, and

A, as G, G¹It is-CH-, and G and G¹ConnectedWhen for double bond, D is connected with GFor singly-bound；

B, when G is-CH- ,-N-, G¹When for-N-, G and G¹ConnectedFor singly-bound, D is connected with GFor list Key；Only when G is-CH-, G¹For-N-, and when D is-N-, G and G¹ConnectedFor singly-bound, D is connected with GFor Double bond；

C, when G is N, G¹When for-CH-, G and G¹ConnectedFor singly-bound, D is connected with GFor singly-bound；

(8) ring T is selected from:

A, five yuan and hexa-atomic aromatic ring or hetero-aromatic ring, described five yuan and hexa-atomic aromatic ring or hetero-aromatic ring can be by R₄Replace, Middle R₄For alkyl, alkenyl, alkynyl, alkoxy, halogen, halogenated alkyl, halogenated alkoxy, hydroxyl, nitro, amino, single alkylamino, Double alkylaminos, alkyl acyl, aminoacyl, alkylamino acyl group, acylamino- ,-CN, aryl or heteroaryl；With

b、C_3-8Cycloalkane or heterocycloalkane, the cycloalkane or heterocycloalkane can be by R₄Replace, wherein R₄For C_1-6Alkane Base, alkenyl, alkynyl, alkoxy, halogen, halogenated alkyl, halogenated alkoxy, hydroxyl, nitro, amino, single alkylamino, double alkane ammonia Base, alkyl acyl, aminoacyl, alkylamino acyl group, acylamino- ,-CN, aryl or heteroaryl.

In some preferred embodiments, the present invention provides the tricyclic fused heterocycle nucleoside phosphoramidate of logical formula (I) Compound, its stereoisomer, pro-drug, pharmaceutically acceptable salt, hydrate, solvate or crystallization, wherein

R₁Selected from H and C_1-6Alkyl；

R₂Selected from C_1-6Alkyl, benzyl；

R₃For there is no, or be selected from C_1-6Alkyl, C_1-6Alkoxy, fluorine, chlorine, bromine, halogenated C_1-6Alkyl, halogenated C_1-6Alcoxyl Base, hydroxyl, nitro, amino, list C_1-6Alkylamino, double C_1-6Alkylamino, acylamino-, ester group ,-CN, C_3-6Naphthenic base, C_3-6Heterocycle alkane Base, aryl, heteroaryl, aralkyl；

D is selected from-(CR_a1R_a2)_mO-、-(CR_a1R_a2)_mS-、-SO-、-SO₂-、-CO-、-N(R_b)-、-N(R_b)-CO-、-C (R_c1R_c2)-, is wherein the R_a1、R_a2Respectively hydrogen or C_1-6Alkyl；The m is 0 or 1；The R_bTo be not present, hydrogen, C_1-6Alkyl, C_1-6Alkyl sulphonyl or C_1-6Alkyl-carbonyl；The R_c1、R_c2Respectively hydrogen, halogen, C_1-6Alkyl, C_2-6Alkenyl Or halogenated C_2-6Alkenyl, or work as R_c1、R_c2It is selected from C_1-3When alkyl, C can be with R_c1、R_c2Constitute C_3-7Naphthenic base；

D¹Selected from oxygen, sulphur ,-SO- ,-SO₂-、-CO-、-N(R_d)-、-(CH)_n, the R_dTo be not present, hydrogen, C_1-6Alkane Base；The n is 0,1 or 2；

G、G¹Respectively-N- or-CH-；And

Ring T is selected from:

(a), at least containing heteroatomic five yuan of hetero-aromatic ring or six-membered Hetero-aromatic or phenyl ring, described five yuan or hexa-atomic Hetero-aromatic ring or phenyl ring can be by R₄Replace, wherein R₄For C_1-6Alkyl, C_1-6Alkoxy, halogen, halogenated C_1-6Alkyl, halogenated C_1-6Alkane Oxygroup, hydroxyl, nitro, amino, list C_1-6Alkylamino, double C_1-6Alkylamino, acylamino- or-CN；With

(b)、C_3-8Cycloalkane or heterocycloalkane, the cycloalkane or heterocycloalkane can be by R₄Replace, wherein R₄For C_1-6 Alkyl, C_1-6Alkoxy, halogen, halogenated C_1-6Alkyl, halogenated C_1-6Alkoxy, hydroxyl, nitro, amino, list C_1-6It is alkylamino, double C_1-6Alkylamino, acylamino- or-CN.

In other preferred embodiments, the present invention provides the tricyclic fused heterocycle nucleosides phosphoramidic acid of logical formula (I) Ester compounds, its stereoisomer, pro-drug, pharmaceutically acceptable salt, hydrate, solvate or crystallization, wherein

R₁Selected from H and C_1-4Alkyl；

R₂Selected from methyl, ethyl, isopropyl, n-propyl, cyclopropyl, cyclopenta, benzyl；

R₃For there is no, or be selected from C_1-4Alkyl, C_1-4Alkoxy, fluorine, chlorine, bromine and phenyl；

D is selected from-(CR_a1R_a2)_mO-、-(CR_a1R_a2)_mS-、-SO-、-SO₂-、-CO-、-N(R_b)-、-N(R_b)-CO-、-C (R_c1R_c2)-, is wherein the R_a1、R_a2Respectively hydrogen, methyl or ethyl；The m is 0 or 1；The R_bTo be not present, Hydrogen, methyl, ethyl, propyl, isopropyl, cyclopropyl, methyl sulphonyl or methyl carbonyl；The R_c1、R_c2Respectively hydrogen, fluorine, Methyl, ethyl, C_2-4Alkenyl or halogenated C_2-4Alkenyl, or work as R_c1、R_c2When being methyl, C and R_c1、R_c2Constitute cyclopropyl；

D¹Selected from oxygen, sulphur ,-SO- ,-SO₂-、-CO-、-N(R_d)-、-(CH)_n, wherein the R_dTo be not present, hydrogen or C_1-4Alkyl；The n is 0 or 1；

G、G¹Respectively N or-CH-；And

Ring T is selected from:

(a), at least containing the heteroatomic five yuan of hetero-aromatic rings of N, O or a S or six-membered Hetero-aromatic or phenyl ring, described five yuan Or six-membered Hetero-aromatic or phenyl ring can be by R₄Replace, wherein R₄For C_1-6Alkyl, C_1-6Alkoxy, halogen, halogenated C_1-6It is alkyl, halogenated C_1-6Alkoxy, hydroxyl, nitro, amino, list C_1-6Alkylamino, double C_1-6Alkylamino, formamido group, acetylamino or-CN；With

(b), pentamethylene, hexamethylene, cycloheptane or at least containing the heteroatomic heterocycle pentane of N, O or a S, heterocycle oneself Alkane, trioxepane.

In some further preferred embodiments, the present invention provides the tricyclic fused heterocycle nucleosides amino of logical formula (I) Phosphate compound, its stereoisomer, pro-drug, pharmaceutically acceptable salt, hydrate, solvate or crystallization, Middle R₁Selected from H and C_1-6Alkyl；R₂Selected from C_1-6Alkyl, benzyl；R₃For there is no, or be selected from C_1-6Alkyl, C_1-6Alkoxy, fluorine, Chlorine, bromine, halogenated C_1-6Alkyl, halogenated C_1-6Alkoxy, hydroxyl, nitro, amino, list C_1-6Alkylamino, double C_1-6Alkylamino ,-CN；D Selected from-(CR_a1R_a2)_mO-、-(CR_a1R_a2)_mS-、-SO-、-SO₂-、-CO-、-N(R_b)-、-N(R_b)-CO-、-(CR_c1R_c2)-, wherein The R_a1、R_a2Respectively hydrogen or C_1-6Alkyl, the m are 0 or 1, the R_bTo be not present, hydrogen, C_1-6Alkyl, C_1-6Alkane Base sulfonyl or C_1-6Alkyl-carbonyl, the R_c1、R_c2Respectively hydrogen, halogen, C_1-6Alkyl, C_2-6Alkenyl or halogenated C_2-6Alkenyl, Or work as R_c1、R_c2It is selected from C_1-3When alkyl, C and R_c1、R_c2Constitute C_3-7Naphthenic base；D¹Selected from oxygen, sulphur ,-SO- ,-SO₂-、-CO-、- NR_d-、-(CH)_n, wherein the R_dTo be not present, hydrogen or C_1-6Alkyl, the n are 0,1 or 2；G,G¹Respectively-N- or- CH-；And ring T is selected from: (a) at least contain heteroatomic five yuan of hetero-aromatic ring or six-membered Hetero-aromatic, phenyl ring, wherein described Five yuan or six-membered Hetero-aromatic or phenyl ring can be by R₄Replace, wherein R₄For C_1-6Alkyl, C_1-6Alkoxy, halogen, halogenated C_1-6Alkyl, Halogenated C_1-6Alkoxy, hydroxyl, nitro, amino, list C_1-6Alkylamino, double C_1-6Alkylamino, acylamino- or-CN；(b) C_3-8Cycloalkanes Hydrocarbon or heterocycloalkane, the cycloalkane or heterocycloalkane can be by R₄Replace, wherein R₄For R₄For C_1-6Alkyl, C_1-6Alkoxy, Halogen, halogenated C_1-6Alkyl, halogenated C_1-6Alkoxy, hydroxyl, nitro, amino, list C_1-6Alkylamino, double C_1-6Alkylamino, acylamino- Or-CN.

In some further preferred embodiments, the present invention provides the tricyclic fused heterocycle nucleosides amino of logical formula (I) Phosphate compound, its stereoisomer, pro-drug, pharmaceutically acceptable salt, hydrate, solvate or crystallization, Middle R₁Selected from H and C_1-4Alkyl；R₂Selected from methyl, ethyl, isopropyl, n-propyl, cyclopropyl, cyclopenta, benzyl；R₃Not deposit , or it is selected from C_1-4Alkyl, C_1-4Alkoxy, fluorine, chlorine, bromine；D is selected from-(CR_a1R_a2)_mO-、-(CR_a1R_a2)_mS-、-SO-、-SO₂-、- CO-、-N(R_b)-、-N(R_b)-CO-、-C(R_c1R_c2)-, is wherein the R_a1、R_a2Respectively hydrogen, methyl or ethyl, the m It is 0 or 1, the R_bTo be not present, hydrogen, methyl, ethyl, propyl, isopropyl, cyclopropyl, methyl sulphonyl or methyl carbonyl, The R_c1、R_c2Respectively hydrogen, fluorine, methyl, ethyl, C_2-4Alkenyl or halogenated C_2-4Alkenyl, or work as R_c1、R_c2When being methyl, C With R_c1、R_c2Constitute cyclopropyl；D¹Selected from oxygen, sulphur ,-SO- ,-SO₂-、-CO-、-N(R_d)-、-(CH)_n, wherein the R_dFor not In the presence of, hydrogen or C_1-4Alkyl, the n are 0 or 1；G,G¹Respectively-N- or-CH-；And ring T is selected from: (a), at least containing one The heteroatomic five yuan of hetero-aromatic rings of a N, O or S or six-membered Hetero-aromatic, phenyl ring, described five yuan or six-membered Hetero-aromatic or phenyl ring can be with By R₄Replace, wherein R₄For C_1-6Alkyl, C_1-6Alkoxy, halogen, halogenated C_1-6Alkyl, halogenated C_1-6Alkoxy, hydroxyl, nitro, ammonia Base, list C_1-6Alkylamino, double C_1-6Alkylamino, formamido group, acetylamino or-CN；(b), pentamethylene, hexamethylene, cycloheptane Or at least contain the heteroatomic heterocycle pentane of N, O or a S, hexamethylene, cycloheptane.

According to the first aspect of the invention, in some preferred embodiments, work as D¹For-(CH)_n, wherein n is 0, i.e., D¹In the absence of, D, G, G¹Phenyl ring in connection constitutes benzo five-membered or hexatomic ring, as shown in logical formula (II)

Wherein

(1)R₁Selected from H and C_1-6Alkyl, the alkyl optionally by one or more alkyl, alkoxy, alkylamino, halogen, Hydroxyl, amino, nitro, cyano, alkyl acyl, aminoacyl, alkylamino acyl group, sulfonyl, sulfinyl, sulfydryl, aryl or Heteroaryl replaces；

(2)R₂Selected from H, C_1-6Alkyl, aminoacyl, aryl and heteroaryl, the alkyl is optionally by one or more alkane Base, alkoxy, halogen, hydroxyl, amino, single alkylamino, double alkylaminos, aryl or heteroaryl replace, such as halogenated C_1-6Alkyl, Benzyl；

(5)G、G¹Respectively-N- or-CH-；

(6)For singly-bound or double bond, two of themIt is singly-bound or oneFor singly-bound, and it is another One is double bond；

(7) ring T is selected from:

A, aromatic ring or heteroaryl, such as can be five yuan and hexa-atomic aromatic ring or hetero-aromatic ring, the aromatic ring or hetero-aromatic ring can be with By R₄Replace, wherein R₄For alkyl, alkoxy, halogen, halogenated alkyl, halogenated alkoxy, hydroxyl, nitro, amino, single alkylamino, Double alkylaminos, acylamino- or-CN；With

In some embodiments, the present invention provides the tricyclic fused heterocycle nucleoside phosphoramidate chemical combination of logical formula (II) Object, its stereoisomer, pro-drug, pharmaceutically acceptable salt, hydrate, solvate or crystallization, wherein

R₁Selected from H and C_1-6Alkyl；

R₂Selected from methyl, ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group, tert-butyl and sec-butyl, cyclopropyl, ring Butyl, cyclopenta, cyclohexyl, benzyl；

R₃For there is no, or be selected from C_1-4Alkyl, C_1-4Alkoxy, fluorine, chlorine, bromine, phenyl；

D is selected from-(CR_a1R_a2)_mO-、-(CR_a1R_a2)_mS-、-SO-、-SO₂-、-CO-、-N(R_b)-、-N(R_b)-CO-、-C (R_c1R_c2)-, is wherein the R_a1、R_a2Respectively hydrogen, methyl or ethyl, the m are 0 or 1, the R_bTo be not present, Hydrogen, methyl, ethyl, propyl, isopropyl, cyclopropyl, methyl sulphonyl or methyl carbonyl, the R_c1、R_c2Respectively hydrogen, fluorine, Chlorine, bromine, methyl, ethyl, C_2-6Alkenyl or halogenated C_2-6Alkenyl, or work as R_c1、R_c2When being methyl, C can be with R_c1、R_c2Constitute cyclopropyl Base；

G、G¹Respectively-N- or-CH-；

For singly-bound or double bond, two of themIt is singly-bound or oneFor singly-bound, and it is another A is double bond,

A, as G, G¹It is-CH-, and G and G¹ConnectedWhen for double bond, D is connected with GFor singly-bound, and

B, when G is-CH- ,-N-, G¹When for-N-, G and G¹ConnectedFor singly-bound, D is connected with GFor list Key；Only when G is-CH-, G¹For-N-, and when D is-N-, G and G¹ConnectedFor singly-bound, D is connected with GFor Double bond；With

Ring T is selected from:

A, at least contain the heteroatomic five yuan of hetero-aromatic rings of N, O or a S or six-membered Hetero-aromatic, phenyl ring, described five yuan or Six-membered Hetero-aromatic or phenyl ring can be by R₄Replace, wherein R₄For C_1-4Alkyl, C_1-4Alkoxy, halogen, halogenated C_1-6It is alkyl, halogenated C_1-4Alkoxy, hydroxyl, nitro, amino, list C_1-4Alkylamino, double C_1-4Alkylamino, acylamino- or-CN；With

B, pentamethylene, hexamethylene, cycloheptane or at least containing the heteroatomic heterocycle pentane of N, O or a S, azacyclohexane, Trioxepane.

In some preferred embodiments, the present invention provides the tricyclic fused heterocycle nucleoside phosphoramidate of logical formula (II) Compound, its stereoisomer, pro-drug, pharmaceutically acceptable salt, hydrate, solvate or crystallization, wherein

R₁For H, C_1-4Alkyl；

R₂Selected from methyl, ethyl, n-propyl, isopropyl, normal-butyl, isobutyl group, tert-butyl and sec-butyl, cyclopropyl, ring Butyl, cyclopenta, benzyl；

R₃For there is no, or be selected from fluorine, chlorine, bromine, phenyl；

D is selected from-(CR_a1R_a2)_mO-、-(CR_a1R_a2)_mS-、-SO-、-SO₂-、-CO-、-N(R_b)-、-N(R_b)-CO-、-C (R_c1R_c2)-, is wherein the R_a1、R_a2Respectively hydrogen, methyl or ethyl, the m are 0 or 1, the R_bTo be not present, Hydrogen, methyl, ethyl, propyl, isopropyl, cyclopropyl, the R_c1、R_c2Respectively hydrogen, fluorine, methyl, ethyl, C_2-6Alkenyl or halogen For C_2-6Alkenyl, or work as R_c1、R_c2Respectively methyl or when ethyl, C can be with R_c1、R_c2Constitute naphthenic base；

G、G¹Respectively N or-CH-；

B, when G is-CH-, N, G¹When for N, G and G¹ConnectedFor singly-bound, D is connected with GFor singly-bound； Only when G is-CH-, G¹For N, and when D is-N-, G and G¹ConnectedFor singly-bound, D is connected with GFor double bond； With

Ring T is selected from:

A, at least contain the heteroatomic five yuan of hetero-aromatic rings of N, O or a S or six-membered Hetero-aromatic, phenyl ring, described five yuan or Six-membered Hetero-aromatic or phenyl ring can be by R₄Replace, wherein R₄For C_1-4Alkyl, C_1-4Alkoxy, halogen, halogenated C_1-6It is alkyl, halogenated C_1-4Alkoxy, hydroxyl, nitro, amino, list C_1-4Alkylamino, double C_1-4Alkylamino, formamido group, acetylamino or-CN；With

B, pentamethylene, hexamethylene, cycloheptane or at least containing the heteroatomic heterocycle pentane of N, O or a S, azacyclohexane, Trioxepane, the naphthenic base or Heterocyclylalkyl can be by R₄Replace, wherein R₄For C_1-4Alkyl, C_1-4It is alkoxy, halogen, halogenated C_1-6Alkyl, halogenated C_1-4Alkoxy, hydroxyl, nitro, amino, list C_1-4Alkylamino, double C_1-4Alkylamino, formamido group, acetyl ammonia Base or-CN.

In other preferred embodiments, the present invention provides the tricyclic fused heterocycle nucleosides phosphoramidic acid of logical formula (II) Ester compounds, its stereoisomer, pro-drug, pharmaceutically acceptable salt, hydrate, solvate or crystallization, wherein

R₁For H, methyl, ethyl；

R₂Selected from methyl, ethyl, isopropyl, n-propyl, cyclopenta, benzyl；

R₃For there is no, or be selected from fluorine, chlorine, bromine；

D is selected from-O- ,-S- ,-SO- ,-SO₂-、-CO-、-N(R_b)-、-N(R_b)-CO-、-C(R_c1R_c2)-, is wherein the R_b To be not present, hydrogen, methyl, ethyl, propyl, isopropyl, the R_c1、R_c2Respectively hydrogen, fluorine, methyl, ethyl, C_2-4Alkenyl, Or work as R_c1、R_c2Respectively methyl or when ethyl, C and R_c1、R_c2Constitute naphthenic base；

G、G¹Respectively-N- or-CH-；

B, when G is-CH- ,-N-, G¹When for N, G and G¹ConnectedFor singly-bound, D is connected with GFor singly-bound； Only when G is-CH-, G¹For-N-, and when D is-N-, G and G¹ConnectedFor singly-bound, D is connected with GIt is double Key；With

Ring T is phenyl ring or hexamethylene, wherein the phenyl ring or hexamethylene can be by R₄Replace, wherein R₄For C_1-4Alkyl, C_1-4 Alkoxy, halogen, halogenated C_1-6Alkyl, halogenated C_1-4Alkoxy, hydroxyl, nitro, amino, list C_1-4Alkylamino, double C_1-4Alkylamino, Formamido group, acetylamino or-CN.

The present invention provides compound in detail below and its non-corresponding isomers, pro-drug, pharmaceutically acceptable salt, Hydrate, solvate:

On the other hand, the present invention provides the preparation method of general formula of the invention (1) compound, and this method includes following step It is rapid:

A) the compound reaction of formula (2) is added under alkaline condition and after phosphorus oxychloride reaction in the compound of formula (1), then Pentafluorophenol is added to react to obtain the compound of formula (3)；With

B) under cryogenic conditions, the compound of formula (3) is reacted with the compound of formula (4), obtains the target compound of formula (5).

Wherein, R₁、R₂、R₃、D、D¹、G、G¹、Ring T has defined above.

The third aspect, the present invention provide pharmaceutical composition, and it includes compounds shown in logical formula (I) of the invention, its solid Isomers, pro-drug, pharmaceutically acceptable salt, hydrate, solvate or crystallization.

In some embodiments, the present invention provides pharmaceutical composition, and it includes compounds shown in logical formula (I), its solid Isomers, pro-drug, pharmaceutically acceptable salt, hydrate, solvate or crystallization, also comprising one selected from following composition Kind or a variety of anti-HCV therapeutic agents: HCV NS3 protease inhibitors, HCV NS5B RNA Dependent RNA polymerase inhibitors, core Glycosides analog, interferon-' alpha ', the interferon of Pegylation, Ribavirin, Levovirin, the pyridine of Wei rummy, TLR7 agonist, TLR9 agonist, cyclophilin inhibitor, α glucosidase inhibitor, NS5A inhibitor and NS3 helicase inhibitors.

It can be by compound shown in logical formula (I) of the invention, its stereoisomer, pro-drug, pharmaceutically acceptable Salt, hydrate, solvate or crystallization are prepared by mixing into drug system with pharmaceutically acceptable carrier, diluent or excipient Agent, to be suitable for oral or parenteral.Medication includes, but are not limited in intradermal, intramuscular, peritonaeum, intravenous, skin Under, intranasal and peroral route.The preparation can be applied by any approach, such as by being transfused or injecting, pass through transepithelial Or the approach application that mucocutaneous (such as oral mucosa or rectum etc.) absorbs.Administration can be whole body or local.Orally The example of application preparation includes solid or liquid dosage form, specifically, including tablet, pill, granula, pulvis, capsule, sugar Slurry, emulsion, suspension etc..The preparation can be prepared by methods known in the art, and include that field of pharmaceutical preparations routinely makes Carrier, diluent or excipient.

Fourth aspect, the present invention provide compound shown in logical formula (I) of the invention, its stereoisomer, pro-drug, medicine Acceptable salt, hydrate, solvate or crystallization or medicine composite for curing flaviviridae infections of the invention on Subject method, compound from logical formula (I) to the subject, stereoisomer, pro-drug, medicine including applying Acceptable salt, hydrate, solvate or crystallization or compound, its stereoisomer, precursor comprising leading to formula (I) on Drug, pharmaceutically acceptable salt, hydrate, solvate or crystallization, to effectively reduce virus described in the subject Virus load amount.In some embodiments, the present invention is provided to treat and/or prevent RNA virus such as flavivirus The method of coe virus infection, including the compound of the present invention, its stereoisomer, salt, hydration are given to individuals in need Object, solvate or crystallization or its pharmaceutical composition.In other embodiments, the present invention, which provides, inhibits RNA virus for example The method of flaviviridae infections, the compound of the present invention, its alloisomerism including making the virus and therapeutically effective amount Body, salt, hydrate, solvate or crystallization or its pharmaceutical composition thereof.

" flaviviridae " refers to any virus of flaviviridae, including infection those of people and non-human animal is viral, Such as flavivirus, pestivirus and Hepatitis C Virus.The compound of the present invention and composition can be particularly useful for the treatment of or prevent Property treatment HCV.

On the other hand, the present invention provides logical formula (I) compound of the invention, its stereoisomer, pro-drug, pharmaceutically Acceptable salt, hydrate, solvate or crystallization are for preventing or treating viral infection, especially flaviviridae infections disease Application, and the application in the drug for preparing preventing/treating viral infectious diseases especially preparing preventing/treating HCV virus infection, as HCV virus hepatitis disease drug in application.

Term definition

Unless otherwise indicated, all technical and scientific terms used herein has and common skill of the art The normally understood identical meaning of art personnel.

As used herein, term " stereoisomer " refers to as produced by the spatially arrangement mode difference of atom in molecule Isomers, including cis-trans-isomer, enantiomter and conformer.All stereoisomers belong to model of the invention It encloses.

As used herein, term " pro-drug " refers to that compound of the present invention ought be applied to mammal offer Any pharmaceutically acceptable form (such as ester, amide compound) of the compound of reactive compound.

As used herein, term " salt " refers to the pharmaceutically acceptable salt that compound of the present invention and acid are formed, The acid for example can be selected from, but be not limited to: phosphoric acid, sulfuric acid, hydrochloric acid, hydrobromic acid, citric acid, maleic acid, malonic acid, almond Acid, succinic acid, fumaric acid, acetic acid, lactic acid, nitric acid, sulfonic acid, p-methyl benzenesulfonic acid, malic acid, Loprazolam or its analog.

As used herein, term " solvate " refers to form the complex of solid-state or liquid by being coordinated with solvent molecule The compounds of this invention form.Hydrate is the special shape of solvate, wherein being coordinated with water.In the scope of the invention Interior, solvate is preferably hydrate.

As used herein, term " crystallization " refers to the various solid forms that compound of the present invention is formed, including crystalline substance It is type, amorphous.

As used herein, term " alkyl " refers to straight chain, branch or cricoid saturated hydrocarbyl, " C_1-6Alkyl " refers to 6 carbon Atom saturated hydrocarbyl below, including linear chain or branched chain C_1-6Alkyl and C_1-6Naphthenic base.For example, suitable C_1-6Alkyl group Including but not limited to methyl, ethyl, n-propyl, isopropyl, cyclopropyl, normal-butyl, isobutyl group, tert-butyl, cyclobutyl, positive penta Base, isopentyl, cyclopenta, cyclohexyl, n-hexyl.The term includes substituted or unsubstituted alkyl, and the alkyl can optional quilt One or more from the following group replaces: alkyl, alkoxy, aryloxy group, alkylamino, arylamino, halogen, hydroxyl, ammonia Base, nitro, cyano, alkyl acyl, aminoacyl, alkylamino acyl group, sulfonyl, sulfinyl, sulfydryl, aryl or heteroaryl.

As used herein, term " alkenyl " refers to the linear chain or branched chain insatiable hunger containing one or more carbon-carbon double bonds (C=C) And alkyl, preferably comprise the alkenyl of 2 to 6 carbon atoms, the alkenyl of further preferably 2 to 4 carbon atoms, most preferably 2 The alkenyl of carbon atom.Term " C_2-6Alkenyl " refers to the unsaturation containing 1 or 2 carbon-carbon double bonds and containing 2 to 6 carbon atoms Alkyl.Term " C_2-4Alkenyl " refers to the unsaturated alkyl containing 1 or 2 carbon-carbon double bonds and containing 2 to 4 carbon atoms, properly Alkenyl group include but is not limited to vinyl, acrylic.Similarly, term " alkynyl " refers to containing one or more carbon carbon three The linear chain or branched chain unsaturated alkyl of key (C ≡ C), preferably comprises the alkynyl of 2 to 6 carbon atoms, further preferably 2 to 4 carbon The alkynyl of atom.

As used herein, term " alkoxy " refers to-O- alkyl, preferably C_1-6Alkoxy, suitable alkoxy base include But be not limited to methoxyl group, ethyoxyl etc..

As used herein, term " halogen " refers to fluorine, chlorine, bromine, iodine.

As used herein, term " halogenated alkyl " refers to the alkyl at least replaced by a halogen atom, preferably halogenated C_1-6 Alkyl.

As used herein, term " alkyl sulphonyl " expression " alkyl-SO₂", preferably C_1-6Alkyl-SO₂, suitable alkyl Sulphonyl groups include but is not limited to methyl sulphonyl, ethylsulfonyl.

As used herein, term " alkyl-carbonyl " expression " C_1-6Alkyl-C (=O)-", suitable alkylcarbonyl-residues include But it is not limited to methyl carbonyl, ethylcarbonyl group.

As used herein, term " aromatic ring " include but is not limited to phenyl ring, naphthalene nucleus, cyclohexyl biphenyl, anthracene nucleus, tetrahydric naphthalene ring, fluorenes ring, Indane ring, sub- cyclohexyl biphenyl and acenaphthene ring, preferably phenyl ring.The term includes replacing and unsubstituted group.The aromatic ring can optional quilt One or more from the following group replaces: alkyl, alkenyl, alkynyl, alkoxy, aryloxy group, halogen, halogenated alkyl, halogenated Alkoxy, hydroxyl, nitro, amino, single alkylamino, double alkylaminos, arylamino, alkyl acyl, aminoacyl, alkylamino acyl Base, acylamino- ,-CN, aryl or heteroaryl.

As used herein, term " hetero-aromatic ring " refers to heteroaromatic containing the 1-4 heteroatomic 3-10 members selected from N, O and S Ring system, preferably C_3-7The heteroaromatic ring system of member, such as thiophene, pyridine, imidazoles, furans, pyrroles, thiazole, 1,2,3- triazoles, 1,2,4- Triazole, 1,2,3- thiadiazoles, oxazole, 1,2,4- oxadiazoles, 1,3,4- oxadiazoles etc., suitable hetero-aromatic ring include but is not limited to Thiophene, pyridine, imidazoles.The term includes replacing and unsubstituted group.The hetero-aromatic ring can be optionally selected from by one or more Group below replaces: alkyl, alkenyl, alkynyl, alkoxy, aryloxy group, halogen, halogenated alkyl, halogenated alkoxy, hydroxyl, nitre Base, amino, single alkylamino, double alkylaminos, arylamino, alkyl acyl, aminoacyl, alkylamino acyl group, acylamino- ,-CN, virtue Base or heteroaryl.

As used herein, term " heterocycloalkane " refers at least containing the heteroatomic cycloalkane of N, O or S, preferably C_3-7 Heterocycloalkane, including but not limited to heterocycle pentane, azacyclohexane, trioxepane.Cycloalkane or heterocycloalkane herein includes Replace and unsubstituted group.The cycloalkane or heterocycloalkane can be optionally substituted with one or more groups selected from the following: Alkyl, alkenyl, alkynyl, alkoxy, aryloxy group, halogen, halogenated alkyl, halogenated alkoxy, hydroxyl, nitro, amino, single alkane ammonia Base, double alkylaminos, arylamino, alkyl acyl, aminoacyl, alkylamino acyl group, acylamino- ,-CN, aryl or heteroaryl.

As used herein, term " acylamino- " is " C_1-6Alkyl-CONH₂", suitable amido groups include but is not limited to Formamido group, acetylamino.

The compound of the present invention contains multiple asymmetric centers, therefore, can be with racemate and racemic mixture, list The form presence of one enantiomter, non-corresponding isomers, non-corresponding isomer mixture and single non-corresponding isomers.

Specific embodiment

The present invention is further elaborated below with reference to embodiment, but the present invention is not limited to these Examples.Except another It is described, the reagent and raw material that the present invention uses are commercially available gained.

Embodiment 1 (2S) -2- (((9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxo -3,4- Dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) propionic acid is different Propyl ester

Step 1, (2S) -2- (((phenyl-pentafluoride oxygroup) (9H- carbazole -4- base oxygroup) phosphoryl) amino) isopropyl propionate Preparation

Add phosphorus oxychloride (0.84g, 0.01mol) in reaction flask, add methylene chloride (DCM, 30mL), -60 DEG C are cooled to, The dichloromethane solution of 4- hydroxycarbazole (1g, 0.01mol) and triethylamine (0.55g, 0.01mol) is added dropwise, drop finishes, room temperature reaction Overnight.This mixed liquor is cooled to -60 DEG C, l-Alanine isopropyl ester hydrochloride is added, triethylamine is added dropwise after stirring 20min The dichloromethane solution of (1.38g, 0.025mol), is added dropwise, and is warming up to -5 DEG C, be added dropwise Pentafluorophenol (1g, 10mmol) and The dichloromethane solution of triethylamine (0.82g, 0.015mol), drop finishes, until being stirred at room temperature.Fully reacting adds water to extract, dry, Concentration, silica gel chromatograph post separation obtain title compound.

Step 2, (2S) -2- (((9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxo -3,4- two Hydrogen pyrimidine -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) propionic acid isopropyl The preparation of ester

It is different that (2S) -2- (((phenyl-pentafluoride oxygroup) (9H- carbazole -4- base oxygroup) phosphoryl) amino) propionic acid is added in reaction flask Propyl ester (26mg, 0.1mmol), sealing, argon gas protection.It injects tetrahydrofuran (THF, 1.5mL), the cooling lower injection tert-butyl of ice bath Magnesium chloride (0.3mL) reacts at room temperature 3h.Under ice bath, the injection fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- (70mg, The solution of THF 0.15mmol), reacts 15h, and reaction terminates.It under ice bath, is quenched with the 2N HCl of 6mL, ethyl acetate extraction It takes, saturated sodium bicarbonate solution washing is dry, is concentrated under reduced pressure, and concentrate obtains title compound through silica gel chromatograph post separation, purifying Object.

¹H NMR(300MHz,DMSO-d₆)δppm:11.46(s,1H),11.43(s,1H),8.19(d,1H),7.49(d, 1H),7.39(t,1H),7.32(t,1H),7.30(t,1H),7.16(d,1H),7.13(t,1H),6.18(t,1H),6.01(t, 1H),5.83(d,1H),5.24(s,1H),4.86(q,1H),4.44-4.48(m,1H),4.32-4.33(m,1H),4.05(s, 1H),4.02(q,1H),3.84-3.90(m,2H),1.24(s,3H),1.20(d,3H),1.13(d,6H)。

ESI-MS m/z:[M+H]⁺=619.

Embodiment 2 (2S) -2- (((9- methyl -9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) Isopropyl propionate

The preparation of step 1,4- hydroxyl -9- methyl -9H- carbazole

In reaction flask, 4- hydroxycarbazole (2g) is dissolved in DMF (l5mL), is slowly added to NaH at 0 DEG C (880mg), then to 30min is stirred at room temperature.It adds MeI (1.5g), 1h, TLC display reaction knot are reacted in continuation at room temperature Beam, ethyl acetate (100mL) dilution, organic phase washed with water, 10%LiCl aqueous solution and saturated common salt washing are dry, remove Organic phase is gone to obtain crude product, crude product obtains title compound, faint yellow solid product through silicagel column (PET:EA=10:1) purifying.

¹H NMR(400MHz,DMSO-d₆)δppm:10.10(s,1H),8.16-8.18(d,1H),7.50-7.53(d, 1H),7.38-7.41(t,1H),7.14-7.26(m,2H),6.99-7.01(d,1H),6.60-6.62(d,1H),3.81(s, 3H)。

LC-MS m/z:[M+H]⁺=198.

Step 2, (2S) -2- (((9- methyl -9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) The preparation of isopropyl propionate

With 4- hydroxyl -9- methyl -9H- carbazole, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and (2' R) title compound can be made with the method for embodiment 1 for raw material in the fluoro- 2'- methyluridine of -2'- deoxidation -2'-.

¹H NMR(400MHz,DMSO-d₆)δppm:11.51(s,1H),8.22-8.24(d,1H),7.61-7.63(m, 1H),7.40-7.51(m,4H),7.17-7.24(m,2H),6.24-6.33(m,1H),5.90-6.05(m,2H),5.15(s, 1H),4.83-4.89(m,1H),4.31-4.48(m,2H),3.93-4.14(m,1H),3.84-3.93(m,5H),1.06-1.22 (m,12H)。

LC-MS m/z:[M+H]⁺=633.

Embodiment 3 (2S) -2- (((9- isopropyl -9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- bis- - 1 (2H)-yl of oxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) ammonia Base) isopropyl propionate

The preparation of step 1,4- hydroxyl -9- isopropyl -9H- carbazole

It in reaction flask, is added 4- hydroxycarbazole (1g, 5.46mmol), THF (20mL) is cooled to -10 DEG C after dissolution, delays Slowly add NaH (0.55g, 14mmol), finish, stir 30 minutes, add 2- N-Propyl Bromide 0.74g (6mmol), finish, until room temperature reaction 3h adds water quenching to go out, and ethyl acetate extraction, dry, concentration, concentrate is prepared chromatogram purification, and PET:EA=10% obtains title Compound, taupe liquid.

Step 2, (2S) -2- (((9- isopropyl -9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) The preparation of isopropyl propionate

With 4- hydroxyl -9- isopropyl -9H- carbazole, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and (2' R) the fluoro- 2'- methyluridine of -2'- deoxidation -2'- is raw material, and with the method for embodiment 1, title compound is made.

¹H NMR(500MHz,DMSO-d₆)δppm:11.46(s,1H),8.20(d,1H),7.73-7.13(m,7H), 6.27-5.67(m,3H),5.14-4.31(m,4H),4.02-3.80(m,4H),1.63(d,6H),1.26-1.22(m,6H), 1.16-1.11(m,6H)。

ESI-MS m/z:[M+H]⁺=661.2.

Embodiment 4 (2S) -2- (((9- ethyl -9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) Isopropyl propionate

The preparation of step 1,4- hydroxyl -9- ethyl -9H- carbazole

In reaction flask, it is added 4- hydroxycarbazole (1g, 5.46mmol), is dissolved in THF (20mL), is cooled to -10 DEG C, adds NaH (0.55g, 14mmol), finishes, and after stirring 30 minutes, adds bromoethane (0.66g, 6mmol), finishes, reacts at room temperature 3h, instead It should terminate, water quenching is added to go out, ethyl acetate extraction is dry, and concentration is prepared chromatogram purification, PET:EA=10% obtains title compound Object, taupe liquid.

Step 2, (2S) -2- (((9- ethyl -- 9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) The preparation of isopropyl propionate

With 4- hydroxyl -9- ethyl -9H- carbazole, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and (2' R) the fluoro- 2'- methyluridine of -2'- deoxidation -2'- is raw material, and with the method for embodiment 1, title compound is made.

¹H NMR(500MHz,DMSO-d₆)δppm:11.47(s,1H),7.61-8.23(m,3H),7.15-7.50(m, 5H),5.96-6.02(m,2H),5.64-5.75(m,2H,),5.16-5.32(m,2H),4.84-4.88(m,1H),4.44- 4.48(m,2H,),4.30-4.37(m,1H),4.05-408(m,1H),3.61-3.65(m,1H),1.21-1.33(m,9H), 1.12-1.17(m,6H)。

ESI-MS m/z:[M+H]⁺=647.2.

Embodiment 5 (2S) -2- (((9- oxo -9H- fluorenes -2- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxo - 3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) third Isopropyl propionate

It is de- with 2- hydroxyl -9-Fluorenone, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and (2 ' R) -2 ' - Oxygen -2 '-fluoro- 2 '-methyluridine is raw material, and with the method for embodiment 1, title compound is made.

¹H NMR(300MHz,DMSO-d₆)δ:11.43(s,1H),7.96(d,1H),7.81(d,1H),7.77(d,1H), 7.59(t,2H),7.41-7.46(m,1H),7.36(t,1H),6.01(d,1H),5.62-5.66(m,2H),5.57(d,1H), 4.82-4.85(m,1H),4.39-4.42(m,1H),4.26-4.29(m,1H),4.02-4.04(m,1H),3.80-3.90(m, 2H),3.62-3.65(m,1H),1.27(d,3H),1.23(d,6H),1.13(dd,3H)。

ESI-MS m/z:[M+Na]⁺=654.2.

Embodiment 6 (2S) -2- (((6,7,8,9- tetrahydro dibenzo [b, d] furans -2- base oxygroup) (((2R, 3R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxy Base) phosphoryl) amino) isopropyl propionate

Steps 1 and 2-hydroxyl-6,7, the preparation of 8,9- tetrahydro dibenzo [b, d] furans

Benzoquinones (5g, 46mmol) is dissolved in toluene, at 0 DEG C be added dropwise 4- (cyclohexyl -1- alkene -1- base) morpholine (8.5g, 51mmol), filtering overnight, filter residue dry 4h after sufficiently being washed with toluene are stirred to react for 0 DEG C.100mL water is added, HCl is added dropwise 18h is stirred at room temperature in (20mL, l5M), and filtering, filter residue is sufficiently washed with water, and after being dissolved with methylene chloride plus water extraction, drying have Machine layer, column purification obtain title compound.

Step 2, (2S) -2- (((6,7,8,9- tetrahydro dibenzo [b, d] furans -2- base oxygroup) (((2R, 3R, 4R, 5R) - 5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) Phosphoryl) amino) isopropyl propionate preparation

With 2- hydroxyl -6,7,8,9- tetrahydro dibenzo [b, d] furans, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, five Fluorophenol and the fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- are raw material, and with the method for embodiment 1, title compound is made.

¹H NMR(300MHz,DMSO-d₆)δppm:11.51(s,1H),7.52(s,1H),7.43(d,1H),7.29(s, 1H),7.04(d,1H),6.02(d,1H),5.97(d,1H),5.84(s,1H),5.49(d,1H),4.37-4.39(m,1H), 4.23-4.24(m,1H),4.01-4.05(m,2H),3.79-3.84(m,2H),2.70(m,2H),2.49(m,2H),1.86- 1.87(m,2H),1.77-1.78(m,2H),1.25(d,3H),1.21(d,3H),1.13(t,6H)。

ESI-MS m/z:[M+H]⁺=624.3.

Embodiment 7 (2S) -2- (((dibenzo [b, d] thiophene -2- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) Isopropyl propionate

The preparation of steps 1 and 2-hydroxyl-dibenzo [b, d] thiophene

The preparation of 1.12- (dibenzo [b, d] thiophene -2- base) -4,4,5,5- tetramethyl -1,3,2- dioxo borine

In dry reaction flask, be added the bromo- dibenzo of 2- [b, d] thiophene (2.64g, 10.0mmol), connection boric acid frequency that Alcohol ester (6.09g, 24.0mmol), potassium acetate (2.94g, 30.0mmol), Isosorbide-5-Nitrae-dioxane (50ml) and [1,1'- bis- (hexichol Base phosphine) ferrocene] palladium chloride (Pd (dppf) Cl₂, 364mg, 0.5mmol).It is heated to 90 DEG C under nitrogen protection, reaction For 24 hours, TLC is detected.Reaction terminates, and reaction system is cooled to room temperature, and water (100mL) is added thereto, is extracted with ethyl acetate (60mL × 3) three times, merge organic phase, washed once with saturated salt solution (60mL).It dries, filters, is concentrated, it is pure through silicagel column Change obtains title compound, yellow oily.

¹H NMR(500MHz,DMSO-d₆) δ ppm:8.61 (s, 1H), 8.25-8.23 (m, 1H), 7.88-7.82 (m, 3H), 7.45-7.43(m,2H),1.39-1.33(m,12H)。

The preparation of 1.22- hydroxyl-dibenzo [b, d] thiophene

In dry reaction flask, compound 2- (dibenzo [b, d] thiophene -2- base)-made from above-mentioned steps 1.1 is added 4,4,5,5- tetramethyls -1,3,2- dioxo borine (2.0g, 6.44mmol), ethyl alcohol (20ml), hydroxylamine hydrochloride (1.34g, 19.32mmol) and sodium hydrate solid (1.03g, 25.76mmol).It is stirred at room temperature after ten minutes, is heated under nitrogen protection For 24 hours, TLC is detected for 60 DEG C of reactions.Reaction system is cooled to room temperature, and water (100mL) is added thereto, is extracted with ethyl acetate (60mL) three times, merges organic phase, then washed once with saturated salt solution (60mL).It dries, filters, is concentrated, through silicagel column Purifying obtains title compound, yellow oily.

¹H NMR(500MHz,DMSO-d₆)δppm:8.06-8.05(m,1H),7.83-7.81(m,1H),7.69-7.37 (m,1H),7.59-7.57(m,1H),7.45-7.41(m,2H),7.01-6.99(m,1H),4.79(brs,1H)。

Step 2, (2S) -2- (((dibenzo [b, d] thiophene -2- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) The preparation of isopropyl propionate

With the 2- hydroxyl of preparation-dibenzo [b, d] thiophene, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol The fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- is raw material, and with the method for embodiment 1, title compound is made.

¹HNMR(500MHz,DMSO)δppm:11.47(s,1H),8.29(d,1H),8.19(s,1H),8.02(m,2H), 7.58-7.56(m,1H),7.54-7.50(m,2H),7.41-7.40(d,1H),6.13-6.03(m,2H),5.85(d,1H), 5.55(d,1H),4.87-4.82(m,1H),4.44-4.38(m,1H),4.30-4.28(m,1H),4.05-4.01(m,1H), 3.90-3.86(m,2H),1.28-1.23(m,6H),1.18-1.12(m,6H)。

ESI-MS m/z:[M+H]⁺=636.0.

Embodiment 8 (2S) -2- (((9- oxo -9H- thioxanthene -2- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- bis- - 1 (2H)-yl of oxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) ammonia Base) isopropyl propionate

The preparation of steps 1 and 2-hydroxyl-9- oxo-9H- thioxanthene

Thiosalicylic acid (9g), phenol (24.7g) put into reaction flask, are then slowly added into the concentrated sulfuric acid (85mL), react 2h is heated at 75 DEG C, TLC monitoring stops reaction.Reaction solution is poured slowly under stiring in 60 DEG C of water, there is solid precipitation, Filtering, solid recrystallize with dichloromethane obtain title compound, light green solid.

¹H NMR(400MHz,DMSO-d₆)δppm:10.24(s,1H),8.49-8.52(m,1H),7.80-7.92(m, 4H),7.53-7.63(m,1H),7.31-7.34(m,1H)。

Step 2, (2S) -2- (((9- oxo -9H- thioxanthene -2- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) The preparation of isopropyl propionate

With 2- hydroxyl -9- oxo -9H- thioxanthene, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and (2' R) the fluoro- 2'- methyluridine of -2'- deoxidation -2'- is raw material, and with the method for embodiment 1, title compound is made.

¹H NMR(400MHz,DMSO-d₆)δppm:11.49(s,1H),8.41-8.44(d,1H),8.24(s,1H), 7.83-7.91(m,3H),7.56-7.76(m,3H),6.19-6.22(t,1H),5.94-5.99(m,2H),5.86(s,1H), 4.75-4.82(m,1H),4.23-4.29(m,2H),3.78-3.82(m,3H),1.11-1.24(m,6H),1.05-1.11(m, 6H)。

LC-MS m/z:[M+H]⁺=664.

Embodiment 9 (2S) -2- (((9- oxo -9H- xanthene -3- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- bis- - 1 (2H)-yl of oxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) ammonia Base) isopropyl propionate

The preparation of step 1,3- hydroxyl -9- oxo -9H- xanthene

The preparation of 1.13- methoxyl group -9- oxo -9H- xanthene

In reaction flask, -4 methoxybenzaldehyde of 2- hydroxyl (1.5g) is dissolved in DMF (40mL), is slowly added at room temperature Enter 1,2- dibromobenzene (4.6g) and potassium carbonate (2.76g), be then heated to 120 DEG C, stir 12h under nitrogen protection, TLC is shown Reaction terminates, and is cooled to room temperature, and is diluted with ethyl acetate (150mL), organic phase washed with water, 10%LiCl aqueous solution and full And brine It, it is dry, organic phase is removed, crude product obtains title compound through silicagel column (PE:EA=10:1) purifying, white Solid product.

¹H NMR(400MHz,DMSO-d₆)δppm:8.16-8.19(q,1H),8.09-8.12(d,1H),7.83-7.88 (m,1H),7.62-7.65(m,1H),7.45-7.50(m,1H),7.17-7.20(d,1H),7.05-7.08(dd,1H),3.94 (s,3H)。

The preparation of 1.23- hydroxyl -9- oxo -9H- xanthene

In reaction flask, by compound 3- methoxyl group -9- oxo -9H- xanthene (300mg) made from above-mentioned steps 1.1 It is dissolved in methylene chloride (5mL), under dry ice acetone bath and nitrogen protection, BBr is added dropwise₃(983mg), stirring are tied until reaction Beam is gone out with 1mL water quenching, and 10mL methylene chloride, organic phase washed with water, saturated common salt water washing is added, and dry methylene chloride subtracts Pressure boils off organic phase and obtains crude product, and title compound, white solid product are obtained after silica gel column purification (PE:EA=5:1).

¹H NMR(400MHz,DMSO-d₆)δppm:11.00(s,1H),8.15-8.17(t,1H),8.04-8.06(d, 1H),7.29(m,1H),7.60-7.63(m,1H),7.45(m,1H),6.88-6.93(m,2H)。

Step 2, (2S) -2- (((9- oxo -9H- xanthene -3- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) The preparation of isopropyl propionate

With 3- hydroxyl -9- oxo -9H- xanthene, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and (2' R) the fluoro- 2'- methyluridine of -2'- deoxidation -2'- is raw material, and with the method for embodiment 1, title compound is made.

¹H NMR(400MHz,DMSO-d₆)δppm:11.51(s,1H),8.18-8.23(m,2H),7.87-7.90(m, 1H),7.66-7.68(d,1H),7.49-7.55(m,3H),7.33-7.35(m,1H),6.33-6.40(m,1H),5.91-6.07 (m,2H),5.57-5.59(d,1H),4.82-4.86(m,1H),4.35-4.44(m,2H),3.88-4.03(m,3H),1.14- 1.29(m,6H),1.10-1.15(m,6H)。

LC-MS m/z:[M+H]⁺=648.

Embodiment 10 (2S) -2- (((9H- carbazole -2- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxo -3,4- Dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) propionic acid is different Propyl ester

It is de- with 2- hydroxyl -9H- carbazole, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and (2'R) -2'- The fluoro- 2'- methyluridine of oxygen -2'- is raw material, and with the method for embodiment 1, title compound is made.It is obtained after purification by HPLC Isomers I, white solid and isomers II, white solid.

HPLC condition: instrument: Shimadzu Corporation prepares HPLC (Preparative liquid chromatography Shimadzu)；Chromatographic column: Benetnoch^TM- C18,20x250mm, 10 μm；Detection wavelength: 254nm；Mobile phase H₂O:MeOH= 30:70。

Corresponding isomers I:

¹H NMR(300MHz,DMSO-d₆)δ:11.48(s,1H),11.27(s,1H),8.06(d,2H),7.60-7.58 (d,1H),7.46-7.45(d,1H),7.36-7.35(d,2H),7.33-7.31(m,1H),7.14-7.13(d,1H),6.08- 6.12(m,2H),5.90(d,1H),5.59(d,1H),4.91-4.82(m,1H),4.67-4.32(m,1H),4.32-4.21(m, 1H),4.05-4.01(m,1H),3.88-3.79(m,2H),2.3(s,3H),1.58(d,3H),1.30-1.20(m,9H)。

ESI-MS m/z:[M+Na]⁺=641.3.

Corresponding isomers II:

¹H NMR(300MHz,DMSO-d₆)δppm:11.48(s,1H),11.26(s,1H),8.01(d,2H),7.61- 7.59(d,1H),7.47-7.45(d,1H),7.36-7.34(d,2H),7.33-7.31(m,1H),7.14-7.13(d,1H), 6.08-6.12(m,2H),5.91(d,1H),5.59(d,1H),4.91-4.82(m,1H),4.67-4.32(m,1H),4.32- 4.21(m,1H),4.05-4.01(m,1H),3.88-3.79(m,2H),2.3(s,3H),1.30-1.20(m,9H),1.58(d, 3H)。

ESI-MS m/z:[M+Na]⁺=641.3.

Embodiment 11 (2S) -2- (((2,3,4,9- tetrahydro -1H- carbazole -6- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2, - 1 (2H)-yl of 4- dioxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphinylidyne Base) amino) isopropyl propionate

Step 1,6- hydroxyl -2,3, the preparation of 4,9-1H- tetrahydro carbazoles

In reaction flask, it is added to methoxyphenyl hydrazine hydrochloride (5g, 29mmol), acetic acid (30mL), cyclohexanone (2.8g, 29mmol), 70 DEG C, Quan Rong are heated to, continues stirring after five minutes, a large amount of yellow solids are precipitated, and TLC is monitored to fully reacting, drop To room temperature, add HBr/H₂O solution 30mL is heated to 80 DEG C, reacts 4 hours, and TLC, which is monitored to raw material, to disappear, and stops reaction, drop To room temperature, ethyl acetate and water is added to extract, organic phase is washed 2 times with saturated sodium bicarbonate solution, dry, concentration, through preparing color It composes column purification (PET:EA=20%), obtains title compound, yellow solid.

Step 2, (2S) -2- (((2,3,4,9- tetrahydro -1H- carbazole -6- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- - 1 (2H)-yl of dioxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) Amino) isopropyl propionate

With 6- hydroxyl -2,3,4,9-1H- tetrahydro carbazole, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and The fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- is raw material, and with the method for embodiment 1, title compound is made.

¹H NMR(500MHz,DMSO-d₆)δppm:11.47(s,1H),10.64(s,1H),7.51(d,1H),7.17- 7.11(m,2H),6.84(d,1H),6.03-5.99(m,1H),5.86-5.80(m,2H),5.50-5.44(m,1H),4.88- 4.83(m,1H),4.36-4.33(m,1H),4.22-4.19(m,1H),4.05-4.00(m,1H),3.84-3.75(m,2H), 2.68-2.54(m,4H),1.81-1.78(m,4H),1.26-1.21(m,6H),1.17-1.14(m,6H)。

ESI-MS m/z:[M+H]⁺=623.0.

Embodiment 12 (2S) -2- (((9H- carbazole -3- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxo -3,4- Dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) propionic acid is different Propyl ester

The preparation of step 1,3- hydroxyl -9H- carbazole

The preparation of 1.13- formoxyl -9- tert-butoxycarbonyl -9H- carbazole

In reaction flask, N-Boc-3- bromine carbazole (3g) is dissolved in THF (10mL), be cooled under nitrogen protection- It 78 DEG C, is added n-BuLi (6.5mL), then reaction solution is added DMF (952mg) thereto, after adding, stops in -78 DEG C of stirring 1h It only cools down, is warmed to room temperature reaction temperature naturally, with the NH of saturation₄Cl solution (1mL) is quenched, and is diluted with EA (20mL), organic It is mutually successively washed with water, saturated common salt, dry organic phase, concentration removes solvent, and crude product is through silica gel chromatography (PE:EA =10:1) obtain title compound, white solid.

The preparation of 1.23- formoxyl oxygroup -9- tert-butoxycarbonyl -9H- carbazole

In reaction flask, 3- formoxyl -9- tert-butoxycarbonyl -9H- carbazole (600mg) is dissolved in THF (10mL), Metachloroperbenzoic acid (m-CPBA, 688mg) is added, 5h is stirred at room temperature in reaction solution, is diluted, is had with methylene chloride (20mL) Machine mutually successively uses water, and saturated sodium bicarbonate aqueous solution is washed, and dry organic phase, concentration removes solvent, obtains title compound, yellow Solid.

The preparation of 1.33- hydroxyl -9- tert-butoxycarbonyl -9H- carbazole

In reaction flask, 3- formoxyl oxygroup -9- tert-butoxycarbonyl -9H- carbazole (670mg) is dissolved in MeOH/H₂O In (7.5mL (volume ratio 2:1)), K is added₂CO₃(890mg), reaction solution stirs 12h in room temperature, dilute with ethyl acetate (20mL) It releases, organic phase washed with water, saturated salt solution washing, dry organic phase, concentration removes solvent, and crude product is through silica gel column chromatography (PET:EA=20:1) purifying obtains title compound, faint yellow solid.

The preparation of 1.43- hydroxyl -9H- carbazole

3- hydroxyl -9- tert-butoxycarbonyl -9H- carbazole (240mg) is dissolved in methylene chloride (4mL), is dripped at 0 DEG C Add trifluoroacetic acid (TFA, 2mL), stir 2h, reaction terminates, reaction solution poured under stiring in suitable ice water, with saturation Sodium bicarbonate solution adjusts pH to neutrality, is extracted with dichloromethane (10mL × 2), organic phase washed with water, saturated common salt washing It washs, dry methylene chloride, decompression boils off organic phase and obtains crude product, obtains through silica gel column purification (PET:EA=3:1) titled Close object, faint yellow solid.

Step 2, (2S) -2- (((9H- carbazole -3- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxo -3,4- two Hydrogen pyrimidine -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) propionic acid isopropyl The preparation of ester

It is de- with 3- hydroxyl -9H- carbazole, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and (2'R) -2'- The fluoro- 2'- methyluridine of oxygen -2'- is raw material, and with the method for embodiment 1, title compound is made.

¹H NMR(400MHz,DMSO-d₆)δppm:11.53(s,1H),11.28(s,1H),7.59-8.06(m,2H), 7.57-7.60(m,1H),7.40-7.47(m,3H),7.16-7.28(m,1H),7.12-7.14(m,1H),6.03-6.06(m, 2H),5.52-5.55(m,1H),4.82-4.86(m,2H),3.86(m,1H),3.85(m,2H),1.21-1.27(m,6H), 1.11-1.14(m,6H)。

LC-MS m/z:[M+H]⁺=619.

Embodiment 13 (2S) -2- (((9- methyl -9H- carbazole -3- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) Isopropyl propionate

The preparation of step 1,3- hydroxyl -9- methyl -9H- carbazole

The preparation of 1.13- methoxyl group -9- methyl -9H- carbazole

In reaction flask, 3- hydroxyl -9H- carbazole (400mg) is dissolved in anhydrous DMF (10mL) at 0 DEG C, iodomethane is added (0.55mL) is kept for 0 DEG C continue to stir 0.5h.It adds sodium hydride (352mg), stirring after five minutes, is warmed to room temperature stirring extremely TLC display reaction terminates.It is diluted with ethyl acetate (50mL), organic phase washed with water, 10%LiCl solution and saturated common salt Water washing, it is dry, organic phase is removed, crude product obtains title compound through silicagel column (PE:EA=10:1) purifying, and canescence is solid Body.

¹H NMR(400MHz,DMSO-d₆)δppm:8.04-8.06(d,1H),7.58-7.59(d,1H),7.11-7.48 (m,5H),3.94(m,3H),3.83(m,3H)。

LC-MS m/z:[M+H]⁺=212.

The preparation of 1.23- hydroxyl -9- methyl -9H- carbazole

In reaction flask, 3- methoxyl group -9- methyl -9H- carbazole (115mg) is dissolved in methylene chloride (5mL), in dry ice Under acetone bath and nitrogen protection, BBr is slowly added dropwise₃(0.1mL), drop finish, and being warmed to room temperature stirring to TLC display reaction terminates.With Water (1mL) is quenched, and is added methylene chloride (10mL), organic phase washed with water, saturated common salt washing, dry methylene chloride, decompression It boils off organic phase and obtains crude product, obtain title compound through column chromatographic purifying separation (PET:EA=5:1), white solid produces Product.

¹H NMR(400MHz,DMSO-d₆)δppm:9.01(s,1H),8.01-8.03(d,1H),7.37-7.51(m,4H), 7.09-7.13(m,1H),6.94-6.97(m,1H),3.80(s,3H)。

Step 2, (2S) -2- (((9- methyl -9H- carbazole -3- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) The preparation of isopropyl propionate

With 3- hydroxyl -9- methyl -9H- carbazole, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and (2' R) the fluoro- 2'- methyluridine of -2'- deoxidation -2'- is raw material, and with the method for embodiment 1, title compound is made.

¹H NMR(400MHz,DMSO-d₆)δppm:11.53(s,1H),7.95-8.13(m,2H),7.46-7.60(m, 4H),7.18-7.37(m,2H),5.92-6.08(m,3H),5.51-5.55(m,1H),4.83-4.88(m,1H),4.24-2.32 (m,2H),4.03-4.06(m,1H),3.81-3.87(m,5H),1.10-1.27(m,12H)。

LC-MS m/z:[M+H]⁺=633.

Embodiment 14 (2S) -2- (((10- methyl -9- oxo-acridan -2- base oxygroup) (((2R, 3R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxy Base) phosphoryl) amino) isopropyl propionate

Steps 1 and 2-hydroxyl-10- methyl-9- oxo-acridan preparation

The preparation of 1.12- (4- Methoxyphenylamino) benzoic acid

In reaction flask, 2- fluobenzoic acid (1.68g) and P-nethoxyaniline (1.23g) are dissolved in THF (l5mL), It is slowly added to lithium amide (460mg) at room temperature, under nitrogen protection, is heated to 50 DEG C of stirring 4h, reaction terminates.It is cooled to room Temperature is quenched with water (1mL), reaction mixture ethyl acetate (50mL) and concentrated hydrochloric acid (2mL) dilution, organic phase washed with water and Saturated common salt water washing, obtains crude product after concentration, be added isopropanol (10mL), and is heated to 65 DEG C and is completely dissolved to solid, It is cooled to room temperature, adds water (10mL), it is cooling under ice bath, title compound, yellow solid is obtained by filtration.

¹H NMR(400MHz,DMSO-d₆)δppm:12.98(s,1H),9.50(s,1H),7.90-7.93(m,1H), 7.37-7.39(m,1H),7.35(m,2H),6.97-7.02(m,3H),6.71-6.74(m,1H),3.81(s,3H)。

LC-MS m/z:[M+H]⁺=244.

The preparation of 1.22- methoxyl group -9- oxo-acridan

In reaction flask, 2- (4- methoxybenzene amido) benzoic acid (800mg) is suspended in POCl₃In (10mL), heating Reflux completes (LC-MS detection) until reaction, and decompression boils off POCl₃；It is added ethyl alcohol in concentrate: 10% hydrochloric acid=8:1 (V: V) solution (10mL), is heated to reflux 1h, is cooled to room temperature, is added water (10mL), there is yellow solid precipitation, filters, obtains title Compound, yellow solid.

¹H NMR(400MHz,DMSO-d₆)δppm:11.77(s,1H),8.22-8.24(d,1H),7.40-7.73(m, 5H),7.22-7.26(m,1H),3.86(s,3H)。

LC-MS m/z:[M+H]⁺=226.

The preparation of 1.32- hydroxyl -9- oxo-acridan

In reaction flask, 2- methoxyl group -9- oxo -9,10- acridan (670mg) is added to 40%HBr aqueous solution It in (13mL), is then heated to reflux for 24 hours, reaction terminates, and is cooled to room temperature, filters, obtains title compound, yellow solid.

¹H NMR(400MHz,DMSO-d₆)δppm:11.64(s,1H),8.18-8.20(m,1H),7.68-7.69(m, 1H),7.45-7.56(m,3H),7.17-7.27(m,2H)。

LC-MS m/z:[M+H]⁺=212.

The preparation of 1.42- methoxyl group -10- methyl -9- oxo-acridan

In reaction flask, 2- hydroxyl -9- oxo -9,10- acridan (211mg) is dissolved in DMF (3mL), in 0 DEG C Under, it is added NaH (160mg), then stirs 30min, add MeI (0.18mL), until room temperature the reaction was continued 30min, reaction knot Beam.It is quenched with water (1mL), mixture is diluted with ethyl acetate (10mL), organic phase washed with water, 10%LiCl aqueous solution and full And brine It, it is concentrated to get crude product after drying, filtering, is obtained through silica gel column chromatography (PE:EA=3:1) purifying titled Close object, yellow solid.

LC-MS m/z:[M+H]⁺=240.

The preparation of 1.52- hydroxyl -10- methyl -9- oxo-acridan

In reaction flask, 2- methoxyl group -10- methyl -9- oxo -9,10- acridan (156mg) is dissolved in dichloromethane Under dry ice acetone bath and nitrogen protection, BBr is slowly added dropwise in alkane (3mL)₃(0.125mL), drop finish, and it is aobvious to TLC to be warmed to room temperature stirring Show that reaction terminates.It is quenched, is added methylene chloride (10mL) with water (1mL), organic phase washed with water, saturated common salt washing are dry Methylene chloride, decompression boil off organic phase and obtain crude product, obtain title compound through column chromatography for separation (PE:EA=5:1) purifying, Yellow solid product.

¹H NMR(400MHz,DMSO-d₆)δppm:9.69(s,1H),8.29-8.31(d,1H),7.67-7.79(m,4H), 7.26-7.34(m,2H),3.91(s,3H)。

LC-MS m/z:[M+H]⁺=212.

Step 2, (2S) -2- (((10- methyl -9- oxo-acridan -2- base oxygroup) (((2R, 3R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxy Base) phosphoryl) amino) and isopropyl propionate preparation

With 2- hydroxyl -10- methyl -9- oxo-acridan, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and the fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- are raw material, and with the method for embodiment 1, title compound is made Object (white solid).

¹H NMR(400MHz,DMSO-d₆)δppm:11.53(s,1H),8.32-8.34(d,1H),8.13-8.14(d, 1H),7.92(m,3H),7.85(m,1H),7.71(m,1H),7.35(m,1H),6.16(t,1H),5.86-6.04(m,2H), 5.53-5.55(d,1H),4.81-4.85(m,1H),4.41(m,1H),4.28(m,1H),4.01(m,1H),3.96(s,3H), 3.80(m,2H),1.22-1.28(m,6H),1.10-1.134(m,6H)。

LC-MS m/z:[M+H]⁺=661.

Embodiment 15 (2S) -2- (((the fluoro- 9H- carbazole -4- base oxygroup of 1-) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) Isopropyl propionate

The preparation of the fluoro- 4- hydroxyl -9H- carbazole of step 1,1-

The preparation of the fluoro- 2- methoxyphenylboronic acid of 1.15-

The bromo- 4- fluoroanisole (5g) of 2- is thrown in reaction flask, is then added anhydrous THF (8OmL), under ice bath, nitrogen protection N-BuLi (20mL) is slowly added dropwise, drips off, reacts 1h, is slow added into trimethylborate (5.5mL), 3h, TCL is stirred at room temperature Monitoring stops reaction.Reaction solution is adjusted to pH value to 1 with 18% hydrochloric acid, and ethyl acetate (200mL) then is added, is saturated chlorination Sodium water solution, organic layer are washed 3 times, and organic phase is merged, and anhydrous sodium sulfate is dry, are concentrated to get title compound, white solid, Crude product can be directly used for reacting in next step.

The preparation of 1.25- fluorine 2- methoxyl group -2'- nitrobiphenyl

By the fluoro- 2- methoxyphenylboronic acid (500mg) of 5-, the bromo- 2- nitrobenzene (591mg) of 1-, [1,1 '-bis- (diphenylphosphines) Ferrocene] palladium chloride dichloromethane complex (215mg), potassium carbonate (811mg) throw in reaction flask, add Isosorbide-5-Nitrae-dioxy Six ring 20mL and water 4mL are heated to 90 DEG C of stirring 1.5h, TCL monitorings under nitrogen protection, stop reaction.Acetic acid is added in reaction solution Ethyl ester (100mL), saturated sodium-chloride water solution (50mL), is extracted with separatory funnel, and organic layer is washed 3 times, merges organic phase, Anhydrous sodium sulfate carrys out drying, is concentrated to get crude product, and crude product is obtained through column chromatographic purifying (petroleum ether: ethyl acetate=100:1-50:1) To title compound, faint yellow solid.

¹H NMR(400MHz,DMSO-d₆)δppm:7.99-8.01(d,1H),7.76-7.79(m,1H),7.63-7.65 (m,1H),7.50-7.53(m,1H),7.23-7.28(m,2H),7.03-7.07(m,1H),3.60(s,3H)。

The preparation of the fluoro- 4- methoxyl group -9H- carbazole of 1.31-

By 5- fluorine 2- -2 '-nitrobiphenyl of methoxyl group (1g), puts into reaction flask, add triphenyl phosphorus (2.1g), 1,2- Dichloro-benzenes (10mL), stirred under nitrogen atmosphere are heated to 190 DEG C overnight.TLC monitoring, stops reaction.Reaction solution is concentrated to get Crude product, crude product obtain title compound, white solid through column chromatography (petroleum ether: ethyl acetate=80:1 to 30:1) purifying.

LC-MS m/z:[M+H]⁺=216.

The preparation of the fluoro- 4- hydroxyl -9H- carbazole of 1.41-

In reaction flask, the fluoro- 4- methoxyl group -9H- carbazole (250mg) of 1- is dissolved in methylene chloride (5mL), in dry ice third Under ketone bath and nitrogen protection, BBr is slowly added dropwise₃(0.22mL), drop finish, and being warmed to room temperature stirring to TLC display reaction terminates.Use water (1mL) is quenched, and is added methylene chloride (20mL), organic phase washed with water, saturated common salt washing, dry methylene chloride, and decompression is steamed It goes organic phase to obtain crude product, obtains title compound, white solid after purification through column chromatography for separation (PET:EA=5:1).

LC-MS m/z:[M+H]⁺=202.

Step 2,2S) -2- (((the fluoro- 9H- carbazole -4- base oxygroup of 1-) (((2R, 3R, 4R, 5R) -5- (dioxo -3 2,4-, 4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) propionic acid The preparation of isopropyl ester

With the fluoro- 4- hydroxyl -9H- carbazole of 1-, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and (2'R) - The fluoro- 2'- methyluridine of 2'- deoxidation -2'- is raw material, and with the method for embodiment 1, title compound, white solid is made.

¹H NMR(400MHz,DMSO-d₆)δppm:11.96-11.96(m,1H),11.60(s,1H),8.26-8.29(d, 1H),7.51-7.61(m,3H),7.15-7.33(m,3H),6.32-6.38(m,1H),5.96-5.98(m,2H),5.34(m, 1H),4.90-4.93(m,1H),4.38-4.54(m,2H),4.11-4.14(m,1H),3.91-3.92(m,2H),1.05-1.14 (m,12H)。

LC-MS m/z:[M+H]⁺=637.

Embodiment 16 (2S) -2- (((the fluoro- 9- methyl -9H- carbazole -4- base oxygroup of 1-) (((2R, 3R, 4R, 5R) -5- (2, - 1 (2H)-yl of 4- dioxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphinylidyne Base) amino) isopropyl propionate

The preparation of the fluoro- 4- hydroxyl -9- methyl -9H- carbazole of step 1,1-

The preparation of the fluoro- 4- methoxyl group -9- methyl -9H- carbazole of 1.11-

By the fluoro- 4- methoxyl group -9H- carbazole (200mg) of 1-, throws in reaction flask, be slowly added to sodium hydride under ice bath stirring (45mg), adds, and the reaction was continued 30 minutes, then adds iodomethane (0.15mL), reacts 1h at room temperature.TLC monitoring, stops Only react.Reaction solution adds water quenching to go out, and ethyl acetate (80mL) then is added, saturated sodium-chloride water solution 50mL, layering, organic layer Washing 3 times merges organic phase, and anhydrous sodium sulfate is dry, is concentrated to get title compound, white solid.

The preparation of the fluoro- 4- hydroxyl -9- methyl -9H- carbazole of 1.21-

In reaction flask, the fluoro- 4- methoxyl group -9- methyl -9H- carbazole (190mg) of 1- is dissolved in methylene chloride (10mL) In, under dry ice acetone bath and nitrogen protection, BBr is added dropwise₃(0.2mL), after -78 DEG C of stirring 2h, then room temperature continues stirring instead 2h is answered, reaction terminates, and is added methylene chloride (10mL), is quenched with the sodium bicarbonate solution that 1mL is saturated, and organic phase washed with water is full And brine It, dry organic layer are concentrated under reduced pressure organic phase and obtain crude product, obtain through silica gel column purification (PE:EA=5:1) Title compound, white solid.

LC-MS m/z:[M+H]⁺=216.

Step 2, (2S) -2- (((the fluoro- 9- methyl -9H- carbazole -4- base oxygroup of 1-) (((2R, 3R, 4R, 5R) -5- (2,4- - 1 (2H)-yl of dioxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) Amino) isopropyl propionate preparation

With the fluoro- 4- hydroxyl -9- methyl -9H- carbazole of 1-, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and The fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- is raw material, and with the method for embodiment 1, title compound is made.

Through HPLC further isolated isomers I (white solid) and isomers II (white solid).

HPLC condition:

Instrument: SHIMADZU LC-6A；Chromatographic column: Shim-pack PREP-ODS (H) (250*20mm, 10um)；Flow velocity: 12mL/min；Detection wavelength: 220/254nm；Mobile phase: A phase: H₂O (0.1%FA) B phase: CH₃OH。

Isomers I:¹H NMR(400MHz,DMSO-d₆)δppm:11.52(s,1H),8.24-8.26(d,1H),7.66- 7.68(d,1H),7.45-7.59(m,2H),7.12-7.30(m,3H),6.28-6.35(m,1H),5.91-6.03(m,2H), 5.21(s,1H),4.83-4.87(t,1H),4.31-4.44(m,2H),4.05(m,4H),3.84-3.89(m,2H),1.18- 1.23(m,6H),1.12-1.24(m,6H)。

LC-MS m/z:[M+H]⁺=651.

Isomers II:¹H NMR(400MHz,DMSO-d₆)δppm:11.53(s,1H),8.23-8.25(d,1H),7.65- 7.67(d,1H),7.5-7.56(m,2H),7.12-7.18(m,3H),6.33-6.37(m,1H),5.97-6.07(m,2H), 5.21-5.27(m,1H),4.73-4.76(t,1H),4.35-4.36(m,2H),4.04-4.09(m,4H),3.85-3.89(m, 2H),1.20-1.23(m,6H),0.99-1.14(m,6H)。

LC-MS m/z:[M+H]⁺=651.

Embodiment 17 (2S) -2- (((9- oxo -9H- xanthene -2- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- bis- - 1 (2H)-yl of oxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) ammonia Base) isopropyl propionate

The preparation of steps 1 and 2-hydroxyl-9- oxo-9H- xanthene

The preparation of 1.12- methoxyl group -9- oxo -9H- xanthene

In reaction flask, 2- nitrobenzaldehyde (1.5g) is dissolved in toluene (10mL), is slowly added at room temperature to first Oxygroup phenol (1.24g), copper chloride (66mg), triphenylphosphine (196mg) and potassium phosphate (4.6g), reaction solution is exposed to air In, it is heated to 110 DEG C of stirrings for 24 hours, TLC display reaction terminates, and is cooled to room temperature, filters, filtrate is dilute with ethyl acetate (150mL) It releases, organic phase water and saturated common salt water washing, it is dry, organic phase is removed, crude product is purified through silicagel column (PE:EA=10:1) To title compound, faint yellow solid product.

¹H NMR(400MHz,DMSO-d₆)δppm:8.17-8.20(m,1H),7.83-7.90(m,1H),7.60-7.65 (m,2H),7.53-7.55(d,1H),7.44-7.48(m,2H),3.87(s,3H)。

The preparation of 1.22- hydroxyl -9- oxo -9H- xanthene

In reaction flask, 2- methoxyl group -9- oxo -9H- xanthene (200mg) is dissolved in methylene chloride (5mL), is done Under ice acetone bath and nitrogen protection, BBr is added dropwise₃(437mg) continues to stir 2h, reaction knot then at room temperature in -78 DEG C of stirring 2h Beam, is added the sodium bicarbonate solution and methylene chloride (10mL) of 1mL saturation, and organic phase washed with water, saturated common salt water washing are done Dry organic layer is concentrated under reduced pressure organic phase and obtains crude product, obtains title compound after silica gel column purification (PET:EA=5:1), Faint yellow solid product.

¹H NMR(400MHz,DMSO-d₆)δppm:10.00(s,1H),8.17-8.19(m,1H),7.83-7.88(m, 1H),7.55-7.66(m,2H),7.44-7.49(m,2H),7.31-7.35(m,1H)。

Step 2, (2S) -2- (((9- oxo -9H- xanthene -2- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) The preparation of isopropyl propionate

With 2- hydroxyl -9- oxo -9H- xanthene, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and (2' R) the fluoro- 2'- methyluridine of -2'- deoxidation -2'- is raw material, with the method for embodiment 1, is made title compound (white solid).

¹H NMR(400MHz,DMSO-d₆)δppm:11.54(s,1H),8.24-8.27(d,1H),7.93-8.05(m, 2H),7.73-7.82(m,3H),7.54-7.84(m,2H),6.27-6.28(m,1H),5.94-6.13(m,2H),5.60-5.62 (m,1H),4.86-4.89(t,1H),4.31-4.49(m,2H),4.07(m,1H),3.89-3.93(m,2H),1.28-1.34 (m,6H),1.15-1.20(m,6H)。

LC-MS m/z:[M+H]⁺=648.

Embodiment 18 (2S) -2- (((9- methyl -2,3,4,9- tetrahydro -1H- carbazole -6- base oxygroup) (((2R, 3R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxy Base) phosphoryl) amino) isopropyl propionate

The preparation of step 1,6- hydroxyl -9- methyl -2,3,4,9-1H- tetrahydro carbazole

The preparation of 1.16- methoxyl group -9- methyl -2,3,4,9-1H- tetrahydro carbazole

In reaction flask, 6- methoxyl group -2,3,4,9-1H- tetrahydro carbazoles (2.5g, 12mmol) and THF (20mL) is added, Be cooled to -10 DEG C after dissolution, be slowly added to NaH (1.5g, 36mmol), finish, stirring 30 minutes after, then plus iodomethane 2.5mL, It finishes, until room temperature reaction 30 minutes, 50 DEG C are heated to, is reacted 1.5 hours, reaction terminates, and is cooled to room temperature, and add water quenching to go out, Ethyl acetate extraction, dry, concentration prepares chromatogram purification, PET:EA=10% obtains title compound, white solid.

The preparation of 1.26- hydroxyl -9- methyl -2,3,4,9-1H- tetrahydro carbazole

In reaction flask, compound 6- methoxyl group -9- methyl -2,3 made from above-mentioned steps 1.1,4,9-1H- tetrahydros are added Carbazole (0.7g, 3.2mmol), 40%HBr aqueous solution and each 10mL of acetic acid are heated to 110 DEG C, react 6 hours, raw material is substantially anti- It should be cooled to room temperature completely, water and ethyl acetate is added to extract, concentration prepares chromatogram purification, PET:EA=10% obtains title Compound, yellow solid.

Step 2, (2S) -2- (((9- methyl -2,3,4,9- tetrahydro -1H- carbazole -6- base oxygroup) (((2R, 3R, 4R, 5R) - 5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) Phosphoryl) amino) isopropyl propionate preparation

With 6- hydroxyl -9- methyl -2,3,4,9-1H- tetrahydro carbazole, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, five Fluorophenol and the fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- are raw material, and with the method for embodiment 1, title compound is made.

¹HNMR(500MHz,DMSO)δppm:11.48(s,1H),7.49-6.90(m,4H),6.04-5.97(m,2H), 5.90-5.81(m,1H),5.49-5.41(m,1H),4.88-4.83(m,1H),4.38-4.33(m,1H),4.22-4.20(m, 1H),4.02-3.99(m,1H),3.82-3.79(m,2H),3.57(s,3H),2.69(m,4H),1.84-1.77(m,4H), 1.27-1.22(m,6H),1.19-1.13(m,6H)。

ESI-MS m/z:[M+Na]⁺=659.2.

Embodiment 19 (2S) -2- (((9- methyl -9H- carbazole -2- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) Isopropyl propionate

The preparation of steps 1 and 2-hydroxyl-9- methyl-9H- carbazole

The preparation of 1.12- methoxyl group -9- methyl -9H- carbazole

By 2- hydroxyl -9H- carbazole (915mg, 5mmol), THF (15mL) is added in reaction flask, under the conditions of 0 DEG C, in batches It is added NaH (130mg, 20mmol), reacts 30min, THF (5ml) solution of iodomethane (568mg, 20mmol) is added, in 30 DEG C react overnight (about 10h).TLC monitors end of reaction, and water quenching is added to go out, and ethyl acetate extracts (50ml × 3), silica gel column purification, Obtain title compound.

The preparation of 1.22- hydroxyl -9- methyl -9H- carbazole

2- methoxyl group -9- methyl -9H- carbazole (730mg) is added in reaction flask, 40%HBr aqueous solution and ice is added Acetic acid, after 105 DEG C of reactions for 24 hours, TLC monitors end of reaction.20ml water is added in reaction solution, is extracted with dichloromethane 3 times, silicon Rubber column gel column purifying.Obtain title compound.

Step 2, (2S) -2- (((9- methyl -9H- carbazole -2- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) The preparation of isopropyl propionate

With 2- hydroxyl -9- methyl -9H- carbazole, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and (2' R) the fluoro- 2'- methyluridine of -2'- deoxidation -2'- is raw material, and with the method for embodiment 1, title compound is made.

¹HNMR(300MHz,DMSO-d6)δppm:11.45(s,1H),8.10-8.11(d,1H),7.95-7.97(d, 1H),7.60-7.61(m,1H),7.59-7.60(m,1H),7.56-7.58(m,1H),7.44-7.45(m,1H),7.36-7.38 (m,1H),7.19-7.22(m,1H),7.07-7.09(m,1H),6.05-6.07(m,2H),5.91(m,1H),5.26(s,1H), 4.83-4.89(m,1H),4.31-4.48(m,1H),3.92-4.14(m,1H),3.84-3.93(m,5H),1.25-1.26(m, 6H),1.11-1.14(m,6H)。

ESI-MS m/z:[M+H]⁺=633.3.

Embodiment 20 (2S) -2- (((dibenzo [b, d] furans -3- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- bis- - 1 (2H)-yl of oxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) ammonia Base) isopropyl propionate

The preparation of step 1,3- hydroxyl-dibenzo [b, d] furans

The preparation of 1.13- amino-dibenzo [b, d] furans

By 3- nitro dibenzofurans (4g), reduced iron powder (5.26g), ammonium chloride (5g) is thrown in reaction flask, and second is added Alcohol (150mL) flows back in 90 DEG C, until TLC monitors raw material fully reacting, stops reaction.It is cooling, it filters, after filtrate decompression concentration, Through column chromatographic isolation and purification, title compound is obtained.

¹H NMR(400MHz,DMSO-d₆)δppm:7.84-7.82(m,1H),7.71-7.69(d,1H),7.52-7.50 (m,1H),7.30-7.25(m,2H),6.75-6.75(d,1H),6.64-6.62(m,1H),5.57(s,2H)。

LC-MS m/z:[M+H]⁺=184.

The preparation of the bromo- dibenzo of 1.23- [b, d] furans

3- amino-dibenzo [b, d] furans (1g, 5.46mmol) is dissolved in acetonitrile (10mL), under condition of ice bath slowly It is added dropwise nitrous acid special butyl ester (731mg, 7.10mmol), is then slowly added into copper bromide (1.46g, 6.55mmol) again, finishes, instead Liquid is answered to be stirred to react 1 hour to 50 DEG C, reaction terminates, and raw material has reacted, and water (1mL) is added to be quenched, and filters, and dichloro is added in filtrate Methane (20mL) is washed with water (40mL × 2), and anhydrous sodium sulfate is dry, and silica gel column chromatography purifies to obtain title compound, white Solid.

¹H NMR(400MHz,DMSO-d₆)δppm:8.16(m,1H),8.11-8.13(d,1H),8.02(s,1H),7.71- 7.73(d,1H),7.56-7.58(m,2H),7.42-7.45(m,1H)。

The preparation of 1.33- formoxyl-dibenzo [b, d] furans

The bromo- dibenzo of 3- [b, d] furans (247mg, 1mmol) is dissolved in anhydrous tetrahydro furan (3mL), slowly in -78 DEG C It is added dropwise n-BuLi (0.6mL), finishes, continue to stir 30 minutes at -78 DEG C of low temperature, DMF (0.1mL) is then slowly added dropwise again, It drips off, is warmed to room temperature stirring 10 minutes, reaction terminates, and aqueous ammonium chloride solution (0.2mL) is added dropwise and is quenched, silica gel column purification is marked Inscribe compound, yellow solid.

¹H NMR(400MHz,DMSO-d₆)δppm:10.15(s,1H),8.38-8.40(d,1H),8.300(m,1H), 8.24(s,1H),7.98-7.98(m,1H),7.79-7.82(d,1H),7.65-7.66(m,1H),7.47-7.49(m,1H)。

The preparation of 1.43- formyloxy-dibenzo [b, d] furans

3- formoxyl-dibenzo [b, d] furans (160mg, 0.81mmol) is dissolved in anhydrous methylene chloride (4mL), Then it is slowly added into metachloroperbenzoic acid (m-CPBA) under room temperature, finishes, reaction solution is stirred at room temperature overnight to anti- It should terminate.Then water (2mL) is added, saturated sodium bicarbonate aqueous solution (4mL), methylene chloride (20mL), extraction, filtration drying, It is concentrated to give title compound, yellow solid.

The preparation of 1.53- hydroxyl-dibenzo [b, d] furans

3- formyloxy-dibenzo [b, d] furans (130mg, 0.6mmol) is dissolved in methanol (3mL), then slowly 5% sodium hydrate aqueous solution (3mL) is added, finishes, in room temperature reaction 30 minutes, reaction terminated, with the salt acid for adjusting pH value of 2N To 2, it is extracted with dichloromethane, washes, anhydrous sodium sulfate is dry, and column chromatography (petroleum ether: ethyl acetate=50:1) obtains title Compound, white solid.

¹H NMR(400MHz,DMSO-d₆)δppm:9.94(s,1H),7.95-7.98(m,1H),7.88-7.90(m,1H), 7.58-7.60(d,1H),7.32-7.34(m,2H),7.01-7.02(d,1H),6.83-6.86(m,1H)。

Step 2, (2S) -2- (((dibenzo [b, d] furans -3- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) The preparation of isopropyl propionate

With 3- hydroxyl-dibenzo [b, d] furans, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and (2' R) the fluoro- 2'- methyluridine of -2'- deoxidation -2'- is raw material, and with the method for embodiment 1, title compound is made.

¹H NMR(400MHz,DMSO-d₆)δppm:11.51(s,1H),8.11-8.15(m,2H),7.68-7.71(d, 1H),7.60-7.61(m,2H),7.51(m,1H),7.39(m,1H),7.28(m,1H),6.17-6.18(m,1H),5.89(m, 1H),5.88(m,1H),5.56-5.59(d,1H),4.81-4.85(m,1H),4.40(m,1H),4.28(m,1H),4.03(m, 1H),3.85-3.88(m,2H),1.23-1.29(m,6H),1.09-1.12(m,6H)。

LC-MS m/z:[M+H]⁺=620.

Embodiment 21 (2S) -2- (((- 2 '-base oxygroup of spiral shell [cyclopropane -1,9 '-fluorenes]) (((2R, 3R, 4R, 5R) -5- (2, - 1 (2H)-yl of 4- dioxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphinylidyne Base) amino) isopropyl propionate

The preparation of steps 1 and 2 '-hydroxyl spiral shell [cyclopropane -1,9 '-fluorenes]

The preparation of the bromo- 9- of 1.12- (2- bromoethyl) -9H- fluorenes

In reaction flask, 2- bromine fluorenes (2g) is dissolved in THF (10mL), LiHMDS (hexamethyl is slowly added in -78 DEG C Two silicon substrate amido lithiums) (9.8mL), 1h is added and stirs, then to 0 DEG C.Said mixture is slowly added into 1,2- dibromo second In THF (10mL) solution of alkane (10.5g), reaction mixture the reaction was continued at 0 DEG C 1h, TLC display reaction terminates, and uses methanol (1mL) is quenched, and removes solvent, and crude product obtains title compound, yellow solid through silica gel column chromatography (PE:EA=100:1) purifying Product.

¹H NMR(400MHz,DMSO-d₆)δppm:7.83-7.91(m,3H),7.57-7.65(m,2H),7.37-7.41 (m,2H),4.16(m,1H),3.40-3.44(m,2H),2.43-2.50(m,2H)。

The preparation of 1.22 '-bromine spiral shells [cyclopropane -1,9 '-fluorenes]

In reaction flask, the bromo- 9- of 2- (2- bromoethyl) -9H- fluorenes (1.1g) is dissolved in DMF (5mL), slowly in 0 DEG C It is added NaH (248mg), then to reacting 12h at room temperature.TLC display reaction terminates, with saturation NH₄Cl (1mL) solution is quenched, EA (20mL) dilution is added, and is successively washed with 10% LiCl solution (5mL × 3), saturated salt solution (5mL), merges organic phase, Anhydrous sodium sulfate dries, filters, and ethyl acetate is removed under reduced pressure, and crude product is marked through silica gel column chromatography (PE:EA=50:1) purifying Inscribe compound, yellow solid.

¹H NMR(400MHz,DMSO-d₆)δppm:7.87-7.95(m,2H),7.51-7.53(m,1H),7.46(s,1H), 7.31-7.38(m,2H),7.19-7.21(m,1H),1.80-1.86(m,2H),1.74-1.79(m,2H)。

The preparation of 1.32 '-formoxyl spiral shells [cyclopropane -1,9 '-fluorenes]

In reaction flask, 2 '-bromine spiral shells [1,9 '-fluorenes of cyclopropane -] (500mg) are dissolved in THF (5mL), -78 DEG C with Under nitrogen protection, it is slowly added to n-BuLi (0.86mL), maintains to react 1h at a temperature of this；Then DMF (0.5mL) is added, stops It only cools down, room temperature is warmed naturally to, and TLC display reaction terminates, with saturation NH₄Cl (1mL) solution is quenched, and is added EA (20mL) Dilution, and successively use 10% LiCl solution (5mL × 2), saturated salt solution (5mL) washing, merging organic phase, anhydrous sodium sulfate It dries, filters, solvent is removed under reduced pressure, crude product silica gel column chromatography (PET:EA=30:1) purifying obtains title compound, yellow Solid.

¹H NMR(400MHz,DMSO-d₆)δppm:10.03(s,1H),8.16-8.18(d,1H),8.06-8.09(m, 1H),7.92-7.94(m,1H),7.766(s,1H),7.41-7.43(m,2H),7.27-7.30(m,1H),1.90-1.94(m, 2H),1.81-1.84(m,2H)。

The preparation of 1.42 '-formyloxy spiral shells [cyclopropane -1,9 '-fluorenes]

In reaction flask, 2 '-formoxyl spiral shells [1,9 '-fluorenes of cyclopropane -] (210mg) are dissolved in DCM (methylene chloride) In (10mL), it is slowly added to m-CPBA (328mg) (metachloroperbenzoic acid) at room temperature, then 3h is reacted at 25 DEG C, until TLC Display reaction terminates.DCM (20mL) dilution is added, and successively with the NaHCO of saturation₃Solution (5mL × 2), saturated salt solution (5mL) washing, merges organic phase, and anhydrous sodium sulfate dries, filters, and DCM is removed under reduced pressure, and obtains title compound, yellow solid, It is directly used in and reacts in next step.

The preparation of 1.52 '-hydroxyl spiral shells [cyclopropane -1,9 '-fluorenes]

In reaction flask, 2 '-formyloxy spiral shells [1,9 '-fluorenes of cyclopropane -] (260mg) are dissolved in MeOH/H₂O(3mL/ LmL in), it is slowly added to K at room temperature₂CO₃(303mg) then reacts 1.5h at 50 DEG C, until TLC display reaction terminates.Decompression Methanol is removed, is diluted with EA (20mL), and successively use H₂O (5mL × 2), saturated salt solution (5mL) are washed, and organic phase is merged, anhydrous Sodium sulphate dries, filters, and EA is removed under reduced pressure, crude product obtains title compound through silica gel column chromatography (PE:EA=15:1) purifying, light Yellow solid.

¹H NMR(400MHz,DMSO-d₆)δppm:9.48(s,1H),7.67-7.74(m,2H),7.08-7.29(m,3H), 6.74-6.77(m,1H),6.52-6.53(m,1H),1.67-1.71(m,2H),1.60-1.63(m,2H)。

Step 2, (2S) -2- (((- 2 '-base oxygroup of spiral shell [cyclopropane -1,9 '-fluorenes]) (((2R, 3R, 4R, 5R) -5- (2,4- - 1 (2H)-yl of dioxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) Amino) isopropyl propionate preparation

With 2 '-hydroxyl spiral shells [cyclopropane -1,9 '-fluorenes], phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and The fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- is raw material, and with the method for embodiment 1, title compound is made.

¹H NMR(400MHz,DMSO-d₆)δppm:11.44(s,1H),7.88-7.91(s,2H),7.57-7.59(m, 1H),7.17-7.36(m,4H),7.07(s,1H),6.07-6.13(m,2H),5.93(br,1H),5.56-5.58(m,1H), 4.83-4.87(m,1H),4.37-4.41(m,1H),4.25-4.27(m,1H),4.0-4.04(m,1H),3.80-3.87(m, 2H),1.69-1.78(m,4H),1.23-1.28(m,6H),1.13-1.15(m,6H)。

LC-MS m/z:[M+H]⁺=644.

Embodiment 22 (2S) -2- (((pyrido [1,2-a] benzimidazole -7- base oxygroup) (((2R, 3R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphorus Acyl group) amino) isopropyl propionate

The preparation of step 1,7- Hydroxy-pyridine simultaneously [1,2a] benzimidazole

The preparation of 1.11- bromine 4- methoxyl group -2- nitrobenzene

4- methoxyl group -2- nitroaniline (1.68g) is dissolved in acetonitrile (30mL), is then slowly added dropwise under ice bath temperature Nitrite tert-butyl (1.34g), then copper bromide (2.67g) is slowly added in reaction flask, reaction solution adds under nitrogen protection Heat is reacted 2 hours to 80 DEG C, and reaction terminates, and is concentrated under reduced pressure, and obtains title compound through silica gel chromatography (PE:EA=20:1) Object, yellow solid.

The preparation of the bromo- 5- aminoanisole of 1.22-

1- bromine 4- methoxyl group -2- nitrobenzene (2.1g) is dissolved in methanol (10mL), iron powder (1.5g) then is added, then to Concentrated hydrochloric acid (2mL) is added in reaction solution, reaction solution flows back after 2h, is cooled to room temperature, and sodium carbonate solid is added until not having bubble It emerges, is extracted with EA, EA layers are washed with saturated common salt, are dried, filtered, and solvent are removed under reduced pressure, crude product is through silica gel column chromatography point Title compound, flaxen grease are obtained from (PE:EA=20:1) purifying.

LC-MS m/z:[M+H]⁺=202.

The preparation of 1.3N- (the bromo- 5- methoxyphenyl of 2-) pyridyl group -2- amine

The bromo- 5- aminoanisole (1.2g) of 2- is dissolved in anhydrous dioxane (6mL), 2- bromopyridine is added (0.68mL), tris(dibenzylideneacetone) dipalladium (Pd2 (dba) 3,100mg), 4,5- bis- diphenylphosphine -9,9- dimethyl oxa-s Anthracene (Xant-phos, 50mg) and cesium carbonate (2.87g), finish, and reaction solution is heated to 100 DEG C under nitrogen protection and is stirred to react 3 Hour, reaction terminates, and solvent is removed under reduced pressure, direct silica gel column chromatography separation (PET:EA=30:1) obtains title compound, light Yellow oily product.

LC-MS m/z:[M+H] +=280.

The preparation of 1.47- methoxv-pyridine simultaneously [1,2a] benzimidazole

N- (the bromo- 5- methoxyphenyl of 2-) pyridyl group -2- amine (1g) is dissolved in anhydrous acetonitrile (10mL), is then added Potassium carbonate (590mg), N, N'- dimethyl-ethylenediamine (315mg) and cuprous iodide (68mg), reaction solution heats under nitrogen protection Overnight to 80 DEG C of reactions, reaction terminates, and filtering filtrate decompression is concentrated, the isolated title of the direct silica gel column chromatography of concentrate Compound, yellow solid.

¹H NMR(400MHz,DMSO-d₆)δppm:9.06-9.08(d,1H),8.23-8.25(d,1H),7.66-7.68 (d,1H),7.56-7.58(m,1H),7.31-7.32(d,1H),7.01-7.05(m,1H),3.91(s,3H)。

LC-MS m/z:[M+H]⁺=199.

The preparation of 1.57- Hydroxy-pyridine simultaneously [1,2a] benzimidazole

By 7- methoxv-pyridine, simultaneously [1,2a] benzimidazole (210mg) is dissolved in methylene chloride (10mL), be cooled to- 78 DEG C, it is then slowly added into Boron tribromide (550mg), is finished, reacted 1 hour at room temperature, after reaction, use ice water It is quenched, methanol (5mL) then is added, is washed with the sodium bicarbonate aqueous solution of saturation, extracted with ethyl acetate (30mL × 3), nothing Aqueous sodium persulfate is dry, is concentrated to get title compound, faint yellow solid.

¹H NMR(400MHz,DMSO-d₆)δppm:9.55(br,1H),8.95-8.97(d,1H),8.07-8.09(d, 1H),7.45-7.57(m,2H),7.05-7.06(d,1H),6.91-6.94(m,1H),6.84-6.86(m,1H)。

LC-MS m/z:[M+H]⁺=185.

Step 2, (2S) -2- (((pyrido [1,2-a] benzimidazole -7- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2, - 1 (2H)-yl of 4- dioxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphinylidyne Base) amino) isopropyl propionate preparation

With 7- Hydroxy-pyridine simultaneously [1,2a] benzimidazole, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol The fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- is raw material, and with the method for embodiment 1, title compound is made.

¹H NMR(400MHz,DMSO-d₆)δppm:11.51(br,1H),9.07-9.09(d,1H),8.30-8.32(d, 1H),7.55-7.68(m,4H),7.22-7.25(m,1H),7.01-7.04(m,1H),5.92-6.14(m,3H),5.51-5.57 (m,1H),4.82-4.85(m,1H),4.28-4.38(m,2H),4.02-4.04(m,1H),3.80-3.89(m,2H),1.19- 1.28(m,6H),1.12-1.15(m,6H)。

LC-MS m/z:[M+H]⁺=620.

Embodiment 23 (2S) -2- (((pyrido [1,2-a] benzimidazole -8- base oxygroup) (((2R, 3R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphorus Acyl group) amino) isopropyl propionate

The preparation of step 1,8- Hydroxy-pyridine simultaneously [1,2a] benzimidazole

The preparation of 1.12- bromine 4- methoxyl group -1- nitrobenzene

5- methoxyl group -2- nitroaniline (3g) is dissolved in acetonitrile (30mL), is then slowly added dropwise under ice bath temperature Nitrite tert-butyl (2.78mL), then copper bromide (4.7g) is slowly added into reaction flask, reaction solution adds under nitrogen protection Heat is reacted 2 hours to 65 DEG C, and reaction terminates, and direct evaporating solvent under reduced pressure obtains title compound through column chromatographic isolation and purification.

The preparation of the bromo- 4- aminoanisole of 1.22-

2- bromine 4- methoxyl group -1- nitrobenzene (4.3g) is dissolved in ethyl alcohol (15mL), iron powder (4g) and chlorination is then added Ammonium (7.8g) is eventually adding water (15mL), and reaction solution is placed in 90 DEG C of 3 hours of reaction, and reaction terminates, and reaction solution is cooled to room Temperature filters out solid, and washs filter cake with methylene chloride, and filtrate is extracted with dichloromethane, saturated common salt water washing, dry, through column Chromatography purifies to obtain title compound, brown oil.

¹H NMR(400MHz,DMSO-d₆)δppm:6.69-6.96(m,1H),6.67-6.74(m,2H),4.08(s,2H), 3.64(s,3H)。

LC-MS m/z:[M+H]⁺=202.

The preparation of 1.3N- (the bromo- 4- methoxyphenyl of 2-) pyridyl group -2- amine

The bromo- 4- aminoanisole (1.2g) of 2- is dissolved in anhydrous dioxane (6mL), 2- bromopyridine is added (0.68mL), tris(dibenzylideneacetone) dipalladium (100mg), 4,5- bis- diphenylphosphine -9,9- xanthphos (50mg) and Cesium carbonate (2.8) g), it finishes, reaction solution is heated to 100 DEG C under nitrogen protection and is stirred to react 3 hours, and reaction terminates, and depressurizes dense Contracting.Residue is directly through the isolated title compound of silicagel column, yellow solid.

¹H NMR(400MHz,DMSO-d₆)δppm:8.13(s,1H),7.99-8.00(m,1H),7.46-7.54(m,1H), 7.22(m,1H),6.93-6.94(m,1H),6.63-6.66(m,2H),3.76(s,3H)。

LC-MS m/z:[M+H]⁺=280.

The preparation of 1.48- methoxv-pyridine simultaneously [1,2a] benzimidazole

N- (the bromo- 4- methoxyphenyl of 2-) pyridyl group -2- amine (1g) is dissolved in anhydrous acetonitrile (10mL), is then added Potassium carbonate (590mg), N, N'- dimethyl-ethylenediamine (315mg) and cuprous iodide (68mg), reaction solution heats under nitrogen protection Overnight to 80 DEG C of reactions, reaction terminates, and filters out solid, filtrate decompression concentration, concentrate is directly through the isolated title of silicagel column Compound, yellow solid.

¹H NMR(400MHz,DMSO-d₆)δppm:8.98-9.00(d,1H),7.90(s,1H),7.68-7.71(d,1H), 7.58-7.60 (m, 1H), 7.42-7.44 (m, 1H), 7.11-7.14 (m, 1H), 6.92-6.95 (m, 1H), 3.88 (s, 3H).

LC-MS m/z:[M+H]⁺=199.

The preparation of 1.58- Hydroxy-pyridine simultaneously [1,2a] benzimidazole

By 8- methoxv-pyridine, simultaneously [1,2a] benzimidazole (100mg) is dissolved in methylene chloride (3mL), is cooled to -78 DEG C, it is then slowly added into Boron tribromide (0.096mL), is finished, reacts 1 hour at room temperature, after reaction, uses ice water It is quenched, methanol (5mL) then is added, is washed with the sodium bicarbonate aqueous solution of saturation, extracted with ethyl acetate (30mL × 2), nothing Aqueous sodium persulfate is dry, is concentrated to get title compound, faint yellow solid.

¹H NMR(400MHz,DMSO-d₆)δppm:9.54(s,1H),8.84-8.86(d,1H),7.60-7.62(d,1H), 7.54-7.55(m,2H),7.38-7.40(m,1H),7.01-7.04(d,1H),6.85-6.88(m,1H)。

LC-MS m/z:[M+H]⁺=185.

With 8- Hydroxy-pyridine simultaneously [1,2a] benzimidazole, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol The fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- is raw material, and with the method for embodiment 1, title compound is made.

¹H NMR(400MHz,DMSO-d₆)δppm:11.50(s,1H),8.98-8.99(d,1H),8.19(s,1H), 7.77-7.80(m,1H),7.64(m,1H),7.54(m,2H),7.40-7.43(m,1H),6.97-6.99(m,1H),6.07- 6.13(m,2H),5.91(m,1H),5.51-5.55(m,1H),4.80-4.84(m,1H),4.41-4.44(m,1H),4.31(m, 1H),3.88-4.02(m,1H),3.82-3.86(m,2H),1.21-1.26(m,6H),1.08-1.11(m,6H)。

LC-MS m/z:[M+H]⁺=620.

Embodiment 24 (2S) -2- (((9- methyl -9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) Ethyl propionate

With the 4- hydroxyl -9- methyl -9H- carbazole of 2 step 1 of embodiment preparation, dichloro oxygen phosphorus, l-Alanine ethyl ester Salt, Pentafluorophenol and the fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- are raw material, and with the method for embodiment 1, targeted is made Close object.

¹H NMR(300MHz,DMSO-d₆)δppm:11.46(s,1H),8.20-8.23(d,1H),7.61-7.63(m, 1H),7.40-7.50(m,4H),7.17-7.24(m,2H),6.24-6.32(m,1H),5.90-6.05(m,2H),5.15(s, 1H),4.83-4.89(m,1H),4.32(m,1H),4.04(m,1H),3.92-3.93(m,2H),3.84-3.92(m,5H), 1.05-1.22 (m, 6H), 1.01-1.02 (m, 3H).

LC-MS m/z:[M+H]⁺=619.

Embodiment 25 (2S) -2- (((9- methyl -9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) Propionic acid ring pentyl ester

The synthesis of step 1, l-Alanine ring ester hydrochloride

It weighs in l-Alanine (8.9g, 100mmol) investment 250mL round-bottomed flask, cyclopentanol 20ml is added, in ice bath item Under part, it is slowly added to thionyl chloride (11.9g, 100mmol), is to slowly warm up to room temperature, overnight, concentration adds for reaction at room temperature Enter isopropyl ether, be beaten under the conditions of -10 DEG C, a large amount of white solids are precipitated, filters, obtain title compound.

Step 2, (2S) -2- (((9- methyl -9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) Benzyl propionate

With the 4- hydroxyl -9- methyl -9H- carbazole of 2 step 1 of embodiment preparation, phosphorus oxychloride, l-Alanine ring pentyl ester hydrochloric acid Salt, Pentafluorophenol and the fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- are raw material, and with the method for embodiment 1, targeted is made Close object.

¹H NMR(300MHz,DMSO-d₆)δppm:11.15(s,1H),8.21-8.24(d,1H),7.59-7.62(m, 1H),7.41-7.50(m,4H),7.16-7.21(m,2H),6.22-6.26(m,1H),6.04-6.18(m,2H),5.19(s, 1H),5.02-5.17(m,1H),4.44-4.45(m,1H),4.42(m,1H),4.01-4.02(m,1H),3.88-3.92(m, 5H), 1.25-1.28 (m, 6H), 1.21-122 (m, 8H).

LC-MS m/z:[M+H]⁺=659.

Embodiment 26 (2S) -2- (((9- methyl -9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) Benzyl propionate

With the 4- hydroxyl -9- methyl -9H- carbazole of 2 step 1 of embodiment preparation, phosphorus oxychloride, l-Alanine benzyl ester hydrochloric acid Salt, Pentafluorophenol and the fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- are raw material, and with the method for embodiment 1, targeted is made Close object.

¹H NMR(300MHz,DMSO-d₆)δppm:11.508(s,1H),8.22-8.25(d,1H),7.59-7.62(m, 1H),7.47-7.50(m,3H),7.38-7.41(m,4H),7.13-7.18(m,3H),6.36-6.40(m,1H),5.90-6.04 (m,2H),4.83-5.15(m,3H),4.33-4.44(m,2H),4.04(m,2H),3.88(m,3H),3.55(m,2H),1.24- 1.26(m,6H)。

LC-MS m/z:[M+H]⁺=681.

Embodiment 27 (2S) -2- (((9- methyl -9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) Propionic acid peopentyl ester

The synthesis of step 1, l-Alanine peopentyl ester hydrochloride

Using l-Alanine and neopentyl alcohol as raw material, with the method for 25 step 1 of embodiment, title compound is made.

Step 2, (2S) -2- (((9- methyl -9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) Propionic acid peopentyl ester

With the 4- hydroxyl -9- methyl -9H- carbazole of 2 step 1 of embodiment preparation, phosphorus oxychloride, l-Alanine peopentyl ester hydrochloric acid Salt, Pentafluorophenol and the fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- are raw material, and with the method for embodiment 1, targeted is made Close object.

¹H NMR(300MHz,DMSO-d₆)δppm:11.48(s,1H),8.22-8.24(d,1H),7.59-7.62(m, 1H),7.45-7.48(m,4H),7.15-7.22(m,2H),6.29-6.33(m,1H),5.91-5.98(m,1H),5.20-5.22 (s,1H),4.46-4.47(m,1H),4.32(m,1H),4.29-4.31(m,1H),4.06-4.12(m,1H),3.96-3.99 (m, 1H), 3.84-3.92 (m, 4H), 3.80-3.87 (m, 2H), 1.24-1.28 (m, 6H), 1.22-1.23 (m, 9H).

LC-MS m/z:[M+H]⁺=661.

Embodiment 28 (2S) -2- (((9,9- dimethyl -9H- fluorenes -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- bis- - 1 (2H)-yl of oxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) ammonia Base) isopropyl propionate

The synthesis of steps 1 and 2 '-methyl biphenyl -2- methyl formate

It weighs 2- methylphenylboronic acid (1g, 7.35mmol), o-iodobenzoic acid methyl esters (1.75g, 6.7mmol), Pd (dppf) Cl₂(0.47g, 0.67mmol), cesium carbonate (2.8g, 20mmol) are thrown in the eggplant-shape bottle of 250mL, and Isosorbide-5-Nitrae-dioxy six is then added Ring 50mL and water 5mL, the lower 90 DEG C of reactions 1.5h of nitrogen protection are added ethyl acetate (100mL) after reaction, are saturated chlorination Sodium water solution (50mL), is extracted with separatory funnel, is washed 3 times, organic phase is then collected, with anhydrous sodium sulfate come drying, mistake Filter, concentration, column chromatographic purifying obtain title compound.

LC-MS m/z:[M+H]⁺=227.

Step 2, the synthesis of 4- methyl -9-Fluorenone

Step 1 gains 2 '-methyl biphenyl -2- carboxylate methyl ester (1g, 4.42mmol) is weighed, is thrown in the eggplant-shape bottle of 100mL In, concentrated sulfuric acid 2ml is then added, 1.5h is stirred at room temperature, stops reaction, reaction solution is poured into ice water, saturated sodium carbonate is added Solution adjusts pH value to neutrality, is added ethyl acetate (100mL), is extracted with separatory funnel, wash 3 times, then collect organic Phase is dried, filtered with anhydrous sodium sulfate, and concentration, column chromatographic purifying obtains title compound.

LC-MS m/z:[M+H]⁺=195.

The synthesis of step 3,4,9,9- trimethyl -9- fluorenes

In the single port bottle of 50ml, argon gas is replaced 3 times, is measured zinc methide toluene solution (30ml, 30mmol), -50 DEG C Under conditions of plus titanium tetrachloride (3.5ml, 60mmol) stir 1 hour, then by step 2 gains 4- methyl -9-Fluorenone (1.94g, It 10mmol) is added in 2ml methylene chloride, is slowly dropped into single port bottle, after fully reacting, pour into ice water and be quenched, acetic acid is added Ethyl ester (500mL), is extracted with separatory funnel, is washed 3 times, is then collected organic phase, dried, filtered with anhydrous sodium sulfate, Concentration, column chromatographic purifying obtain title compound.

LC-MS m/z:[M+H]⁺=209.

Step 4,9,9- dimethyl -4- methylol -9H- fluorenes

In the single port bottle of 100mL, 4,9,9- trimethyl -9- fluorenes (1.1g, 5mmol) of step 3 gains, four chlorinations is added Acetic acid is added in carbon 5ml, NBS (1g, 5.6mmol) and azodiisobutyronitrile (AIBN, 700mg, 4.2mmol), heated overnight at reflux Ethyl ester (500mL), is extracted with separatory funnel, is washed 3 times, is then collected organic phase, with anhydrous sodium sulfate come drying, be concentrated to give To crude product.By in the single port bottle of gained crude product investment 250mL, potassium carbonate (910mg, 6.6mmol), water 20ml and Isosorbide-5-Nitrae-two is added Six ring 50ml of oxygen is heated to 90 DEG C, reacts 3 hours, fully reacting, is added ethyl acetate (500mL), is extracted with separatory funnel It takes, washes 3 times, then collect organic phase, dried, filtered with anhydrous sodium sulfate, be concentrated, column chromatographic purifying obtains title compound Object.

LC-MS m/z:[M+H]⁺=225.

Step 5,9,9- dimethyl -4- formoxyl -9H- fluorenes

By step 4 gains 9,9- dimethyl -4- methylol -9H- fluorenes (400mg, 1.7mmol) puts into the single port bottle of 50mL In, methylene chloride 15ml is added, adds Pyridinium chlorochromate on silica gel (PCC, 1.2g, 6mmol) under condition of ice bath, after fully reacting, is added Ethyl acetate (200mL), is extracted with separatory funnel, is washed 3 times, organic phase is then collected, with anhydrous sodium sulfate come drying, mistake Filter, concentration, column chromatographic purifying obtain title compound.

LC-MS m/z:[M+H]⁺=223.

The synthesis of step 6,4- hydroxyl -9,9- dimethyl -9- fluorenes

By step 5 gains 9,9- dimethyl -4- formoxyl -9H- fluorenes (200mg, 0.95mmol) puts into the single port of 50mL In bottle, methylene chloride 15ml is added, under ice bath, adds metachloroperbenzoic acid (mCPBA, 500mg, 3mmol), is warmed to room temperature, room Temperature is stirred overnight, and after fully reacting, is added ethyl acetate (200mL), is extracted with separatory funnel, the thio sulphur of organic addition 10% Acid sodium aqueous solution is washed 3 times, is dried, filtered with anhydrous sodium sulfate, after concentrate drying, addition 15ml methanol, KOH (56mg, 1mmol), normal-temperature reaction after fully reacting, is added ethyl acetate (200mL), is extracted with separatory funnel, washing 3 times, then Organic phase is collected, is dried, filtered with anhydrous sodium sulfate, is concentrated, column chromatographic purifying obtains title compound.

LC-MS m/z:[M+H]⁺=211.

Step 7, (2S) -2- (((9,9- dimethyl -9H- fluorenes -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) Isopropyl propionate

With step 6 gains 4- hydroxyl -9,9- dimethyl -9- fluorenes, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, five Fluorophenol and the fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- are raw material, and with the method for embodiment 1, target compound is made.

¹H NMR(300MHz,DMSO-d₆)δppm:11.46(s,1H),8.20-8.23(d,1H),7.66-7.69(m, 1H),7.38-7.47(m,4H),7.19-7.21(m,2H),6.23-6.27(m,1H),6.02-6.19(m,1H),5.86-5.95 (m,1H),5.08(s,1H),4.88-4.90(m,1H),4.45-47(m,1H),4.30-41(m,1H),4.28-4.31(m, 1H),3.92-3.94(m,2H),1.05-1.22(m,18H)。

LC-MS m/z:[M+H]⁺=659.

Embodiment 29 (2S) -2- (((9- cyclopropyl -9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- bis- - 1 (2H)-yl of oxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) ammonia Base) isopropyl propionate

The synthesis of step 1,4- methoxyl group -9H- carbazole

It weighs 4- hydroxycarbazole (9.1g, 50mmol), potassium carbonate (6.9g, 50mmol) is put into the three-necked bottle of 500ml, is added Enter 250ml acetone, under condition of ice bath, be slowly added to dimethyl suflfate (4.8ml, 50mmol), reacts 2h, reaction knot at room temperature Shu Hou, is added ethyl acetate (100mL) dilution, organic phase washed with water, and saturated common salt washing is dried, filtered, is concentrated, column layer Analysis purifies to obtain title compound.

LC-MS m/z:[M+H]⁺=198.

The synthesis of step 2,9- cyclopropyl -4- methoxyl group -9H- carbazole

Weigh step 1 gains 4- methoxyl group -9H- carbazole (3.96g, 20mmol), copper acetate (4.0g, 20mmol), ring Propyl boric acid (3.44g, 40mmol) and DMAP (7.32g, 60mmol) investment 500ml eggplant-shape bottle in, be added 200ml toluene and After fully reacting, it is dilute that ethyl acetate (500mL) is added in 20ml bis- (trimethyl silicon substrate) Sodamide (NaHMDS), 95 DEG C of reaction 4h It releases, organic phase washed with water, saturated common salt washing is dried, filtered, is concentrated, and column chromatographic purifying obtains title compound.LC-MS m/ z:[M+H]⁺=238.

The synthesis of step 3,9- cyclopropyl -4- hydroxyl -9H- carbazole

Weigh the eggplant shape of step 2 gains 9- cyclopropyl -4- methoxyl group -9H- carbazole (1.19g, 5mmol) investment 250ml In bottle, 100ml methylene chloride is added and is slowly added to BBr under the conditions of -50 DEG C₃(7ml, 7.5mmol), finishes, -20 DEG C of reactions 4h after fully reacting, is slowly added to 200ml water, ethyl acetate (500mL) extraction, organic phase washed with water, saturated salt solution It washes, dries, filters, be concentrated, column chromatographic purifying obtains title compound.

LC-MS m/z:[M+H]⁺=224.

Step 4, (2S) -2- (((9- cyclopropyl -9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) The preparation of isopropyl propionate

With step 3 gains 9- cyclopropyl -4- hydroxyl -9H- carbazole, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, five Fluorophenol and the fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- are raw material, and with the method for embodiment 1, target compound is made.

¹H NMR(300MHz,DMSO-d₆)δppm:11.45(s,1H),7.97-7.98(d,1H),7.53-7.56(m, 2H),7.36-7.38(m,2H),7.27-7.30(m,3H),6.20-6.22(m,1H),6.18-6.20(m,1H),6.00-6.05 (m,1H),5.31(s,1H),4.86-4.89(m,1H),4.45-4.47(m,1H),4.32-4.33(m,1H),4.29-4.31 (m, 1H), 3.87-3.89 (m, 2H), 2.42 (m, 1H), 1.42-1.43 (m, 6H), 1.32 (m, 2H), 1.28 (m, 2H), 1.23- 1.24(m,6H)。

LC-MS m/z:[M+H]⁺=659.

Embodiment 30 (2S) -2- (((dibenzo [b, d] furans -1- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- bis- - 1 (2H)-yl of oxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) ammonia Base) isopropyl propionate

The preparation of step 1,3,4,6,7,8,9- hexahydro dibenzo [b, d] furans -1 (2H) -one

In the there-necked flask of 250mL, addition 1, hydroresorcinol (11g, 0.1mol), cyclohexanone (10g, 0.1mol), two Toluene 100ml, p-methyl benzenesulfonic acid 0.45g (0.01mol), bonus point hydrophone, 170 DEG C of reflux 5h after fully reacting, are cooled to room Temperature, concentration, column chromatographic purifying obtain title compound.

The preparation of step 2,1- hydroxyl dibenzo [b, d] furans

In the single port bottle of 250mL, step 1 gains 3,4,6,7,8,9- hexahydro dibenzo [b, d] furans -1 is added (2H) -one 5.2g (0.027mmol), cymene 80mL, 10%Pa/C3g, under nitrogen protection, 170 DEG C of reflux 18 are small When, after fully reacting, cool down, filter, concentration, column chromatographic purifying obtains title compound.

Step 3, (2S) -2- (((dibenzo [b, d] furans -1- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) The preparation of isopropyl propionate

With step 2 gains 1- hydroxyl dibenzo [b, d] furans, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, five fluorine Phenol and the fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- are raw material, and with the method for embodiment 1, target compound is made.

¹HNMR((300MHz,DMSO-d₆)δppm:11.49(s,1H),8.43(m,1H),8.16(d,1H),7.74(d, 1H),7.53-7.38(m,5H),6.34(m,1H),6.05(m,1H),5.89(m,1H),5.37(m,1H),5.19(m,1H),4, 84(m,1H),4.45(m,1H),4.04(m,1H),3.91(m,2H),1,26-1.10(m,12H)。

LC-MS m/z:[M+H]⁺=620.

Embodiment 31 (2S) -2- (((5- methyl -6- oxo -5,6- dihydrophenanthridine -10- base oxygroup) (((2R, 3R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxy Base) phosphoryl) amino) isopropyl propionate

The preparation of step 1,3- hydroxy-n-(2- iodophenyl) benzamide

M-hydroxybenzoic acid (1g, 7.25mmol) is dissolved in 20ML n,N-dimethylacetamide, is added dropwise two at 0-4 DEG C Chlorine sulfoxide (1.3g, 10.9mmol), finishes, and after reacting at room temperature 0.5h, adjacent Iodoaniline (1.6g, 7.25mmol) is added dropwise, finishes, room Temperature is lower the reaction was continued 3h, after fully reacting, adds water and ethyl acetate to extract, dries, filters, be concentrated, column chromatographic purifying obtains title Compound.

The preparation of step 2,3- methoxyl group-N- (2- iodophenyl)-N-methyl-benzamide

In the single port bottle of 100mL, addition step 1 gains 3- hydroxy-n-(2- iodophenyl) benzamide (0.5g, It 1.47mmol) is dissolved in DMF, under ice bath, adds NaH (0.2g, 4.42mmol), after stirring 0.5h, add dimethyl suflfate 1.33g (10.5mml), finishes, room temperature reaction 2h, fully reacting, and under ice bath plus water quenching is gone out, and ethyl acetate and water extraction dry, filter, Concentration, column chromatographic purifying obtain title compound.

The preparation of step 3, -6 (5H) -one of 10- methoxyl group -5- methyl phenanthridines

In the single port bottle of 100mL, add step 2 gains 3- methoxyl group-N- (2- iodophenyl)-N-methyl-benzamide (0.5g, 1.36mmol), is dissolved in DMF20ml, adds Pd (OAc)₂(50mg, 0.22mmol), PPh₃(70mg, 0.267mmol), K₂CO₃(0.4g, 2.9mmol), nitrogen are replaced three times, and 160 DEG C of reflux 3h cool down after fully reacting, and water and ethyl acetate is added to extract It takes, dries, filters, be concentrated, column chromatographic purifying obtains title compound.

The preparation of step 4, -6 (5H) -one of 10- hydroxy-5-methyl base phenanthridines

In the single port bottle of 100mL, add -6 (5H) -one of step 3 gains 10- methoxyl group -5- methyl phenanthridines (0.15g, 0.6mmol), HBr/H₂O (40%, 5ml), acetic acid 5mL, 110 DEG C of reactions after fully reacting, add water and ethyl acetate to extract, do It is dry, it filters, concentration, column chromatographic purifying obtains title compound.

Step 5, (2S) -2- (((5- methyl -6- oxo -5,6- dihydrophenanthridine -10- base oxygroup) (((2R, 3R, 4R, 5R) - 5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) Phosphoryl) amino) isopropyl propionate

With -6 (5H) -one of step 4 gains 10- hydroxy-5-methyl base phenanthridines, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride Salt, Pentafluorophenol and the fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- are raw material, and with the method for embodiment 1, targeted is made Close object.

¹H NMR(500MHz,DMSO-d₆) δ ppm:11.47 (s, 1H), 9.04 (d, 1H), 8.30 (d, 1H), 7.8-7.3 (m,6H),6.39-6.30(m,1H),5.80-6.06(m,2H),5.20(m,1H),4.80(m,2H),4.40-4.25(m,2H), 3.91(m,2H),3.78(s,3H),1.05-1.25(m,12H)。

LC-MS m/z:[M+H] +=661.

Embodiment 32 (2S) -2- (((9- methyl -9H- carbazole -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) Methyl propionate

With 2 step 1 gains 4- hydroxyl -9- methyl -9H- carbazole of embodiment, phosphorus oxychloride, l-Alanine methyl esters hydrochloric acid Salt, Pentafluorophenol and the fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- are raw material, and with the method for embodiment 1, targeted is made Close object.

¹H NMR(300MHz,DMSO-d₆)δppm:11.47(s,1H),8.23(d,1H),7.61-7.63(d,1H), 7.40-7.50(m,4H),7.17-7.24(m,2H),6.24-6.32(m,1H),5.90-6.05(m,2H),5.15(s,1H), 4.83-4.89(m,1H),4.32(m,1H),4.04(m,1H),3.92-3.93(m,5H),3.60(s,3H),1.05-1.25(m, 6H)。

LC-MS m/z:[M+H]⁺=605.

Embodiment 33 (2S) -2- (((9H- fluorenes -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxo -3,4- two Hydrogen pyrimidine -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) propionic acid isopropyl Ester

The preparation of step 1,4- hydroxyl -9H- fluorenes -9- ketone

Using 28 step 2 gains 4- methyl of embodiment -9-Fluorenone as raw material, with the method for 28 step 3 of embodiment, 4,5 and 6 Title compound is made.

The preparation of step 2,4- hydroxyl -9H- fluorenes

In the single port bottle of 100mL, it is added step 1 gains 4- hydroxyl -9H- fluorenes -9- ketone (200mg, 1.02mmol), it is molten In methanol 20ml, add 10% palladium carbon 20mg, hydrogen is divided into lower room temperature reaction overnight, after fully reacting, filters, and concentration obtains title Compound.

Step 3, (2S) -2- (((9H- fluorenes -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxo -3,4- dihydro Pyrimidine -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) methyl propionate

With step 2 gains 4- hydroxyl -9H- fluorenes, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and (2' R) the fluoro- 2'- methyluridine of -2'- deoxidation -2'- is raw material, and with the method for embodiment 1, target compound is made.

¹H NMR(300MHz,DMSO-d₆)δppm:11.46(s,1H),8.23(d,1H),7.61-7.20(m,7H), 6.24-6.32(m,1H),5.85-6.05(m,2H),5.20(s,1H),4.83(m,1H),4.30(m,1H),4.14(m,1H), 3.90(m,1H),3.80(d,2H),3.62(m,2H),1.05-1.25(m,12H)。

LC-MS m/z:[M+H] +=618.2.

Embodiment 34 (2S) -2- (((9- oxo -9H- fluorenes -4- base oxygroup) (((2R, 3R, 4R, 5R) -5- (2,4- dioxy Generation -3,4- dihydro-pyrimidin -1 (2H)-yl) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) phosphoryl) amino) Isopropyl propionate

With 33 step 1 gains 4- hydroxyl -9H- fluorenes -9- ketone of embodiment, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride, Pentafluorophenol and the fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- are raw material, and with the method for embodiment 1, target chemical combination is made Object.

¹H NMR(300MHz,DMSO-d₆)δppm:11.46(s,1H),8.23(d,1H),7.61-7.20(m,7H), 6.24-6.32(m,1H),5.85-6.05(m,2H),5.20(s,1H),4.83(m,1H),4.30(m,1H),4.14(m,1H), 3.90(m,1H),3.80(d,2H),1.05-1.25(m,12H)。

LC-MS m/z:[M+H]⁺=632.

Embodiment 35 (2S) -2- (((5- methyl -6- oxo -5,6- dihydrophenanthridine -1- base oxygroup) (((2R, 3R, 4R, 5R) -5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) the fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxy Base) phosphoryl) amino) isopropyl propionate

The preparation of step 1, N- (3- hydroxy phenyl) -2- iodobenzamide

It in the single port bottle of 500ml, is added m-aminophenol 5g (45.9mmol), is dissolved in 80ml THF, adjacent iodine is added Benzoic acid 14g (56.5mmol), EDCI13g (67.8mmol) and DMAP1.3g (10.6mmol), are stirred overnight at room temperature, have reacted Quan Hou, concentration are added ethyl acetate and water extraction, dry, filter, be concentrated, and column chromatographic purifying obtains title compound.

The preparation of step 2, -6 (5H) -one of 1- hydroxy-5-methyl base phenanthridines

Using step 1 gains N- (3- hydroxy phenyl) -2- iodobenzamide as raw material, according to 31 step 2 of embodiment, 3 and 4 Method be made title compound.

Step 3, (2S) -2- (((5- methyl -6- oxo -5,6- dihydrophenanthridine -1- base oxygroup) (((2R, 3R, 4R, 5R) - 5- (- 1 (2H)-yl of 2,4- dioxo -3,4- dihydro-pyrimidin) fluoro- 3- hydroxy-4-methyl tetrahydrofuran -2- base of -4-) methoxyl group) Phosphoryl) amino) isopropyl propionate

With -6 (5H) -one of step 2 gains 1- hydroxy-5-methyl base phenanthridines, phosphorus oxychloride, l-Alanine isopropyl ester hydrochloride Salt, Pentafluorophenol and the fluoro- 2'- methyluridine of (2'R) -2'- deoxidation -2'- are raw material, and with the method for embodiment 1, targeted is made Close object.

¹H NMR(300MHz,DMSO-d₆)δppm:11.50(s,1H),9.0(d,1H)8.23(d,1H),7.8-7.20(m, 5H),6.40-6.30(m,1H),5.82-6.06(m,2H),5.20(m,1H),4.80(m,2H),4.40-4.25(m,2H), 4.12(m,1H),3.91(m,1H),3.80(m,1H),3.65(s,3H),1.05-1.25(m,12H)。

LC-MS m/z:[M+H] +=661.

1 pharmacology activity research of experimental example:

1. experimental material

1.1 reagents:

1. reagent list of table

Reagent name	Supplier
		DMEM cell culture medium	Invitrogen
Fetal calf serum (FBS)	Gibco
		L-Glutamine	Invitrogen
Pen .- Strep solution	Invitrogen
		DPBS/Modified	Hyclone
Trypsase/EDTA	Invitrogen
		Dimethyl sulfoxide (DMSO)	Sigma
Bright-Glo	Promega
		Cell grows fluorescence titration detection reagent	Promega

1.2Huh71b cell line:

Huh71b cell line is provided by Shanghai Yaoming Kangde New Medicine Development Co., Ltd, for comprising having stable fluorescein The Huh7 cell line of the HCV1b replicon of enzyme (Luc) report.Its by gene recombination technology by HCV nonstructural protein gene, Neo (G418 resistance) and luciferase reporter gene are cloned into pBR vector construction.Then the carrier of HCV replicon will be carried It is transfected into huh7 cell, by G418 resistance screening, HCV replicon can stablize duplication and GAP-associated protein GAP and luciferase are in huh7 Cell inner stablity expression.The cell model is screened for HCV-Ab IgG Compound ira vitro.By the expression for checking luciferase Measure the activity of the HCV-Ab IgG of compound.Referring to Lohmann V, et al.1999.Replication of subgenomic hepatitis C virus RNAs in a hepatoma cell line.Science.285(5424):110-113.

1.3 positive control drugs:

Comparison medicine structure used in experimental example of the present invention are as follows:It is The compound of WO2008/121634 (PCT/US2008/058183) embodiment 25, i.e. (S) -2- { [(2R, 3R, 4R, 5R) -5- (2,4- dioxo -3,4- dihydro -2H- pyrimidine -1- base) fluoro- 3- hydroxy-4-methyl of -4--tetrahydro-furan -2- ylmethoxy] - Phenoxy group-phosphoryl amino }-isopropyl propionate ((S) -2- { [(2R, 3R, 4R, 5R) -5- (2,4-dioxo-3,4-dihydro- 2H-pyrimidin-1-yl)-4-fluoro-3-hydroxy-4-me thyl-tetrahydro-furan-2- ylmethoxy]-phenoxy-phosphorylamino}-propionic acid isopropyl ester)。

The compound referring to J.Org.chem, method described in 2011,76,8311-8319 be made and by hydrogen compose with Mass Spectrometric Identification.

2. experimental procedure:

2.1 compounds prepare: with the full-automatic microwell plate pretreatment system of POD810 (LabCyte company, the U.S.) will more than The compound of the present invention of embodiment preparation is added in orifice plate, and it is 10 μM that compound, which originates final concentration, and each compound is done Duplicate hole, 3 times of dilutions, 10 points, DMSO final concentration 0.5%；

2.2 cells prepare: kind Huh71b cell to 96 orifice plates respectively, 125 μ l systems, and 8 × 10³A cells/well, 37 DEG C, 5%CO₂Incubator culture cell 72 hours；

2.3 cytoactive detections: every hole add 30 μ l cells grow fluorescence titration detection reagent, 37 DEG C, 5%CO₂Incubator Culture cell 1 hour, detects fluorescence signal value on spectrophotometer, the data obtained is calculated for Compound Cytotoxicity；

2.4Bright-Glo detection: every hole adds 100 μ l luciferase luminous substrate Bright-Glo, with chemistry in 5 minutes Luminescent detection system EnVision (PerkinElmer company, the U.S.) detects fluorescence signal value, and the data obtained is imitated for compound Power calculates.

2.5 data processings: the data obtained is converted into cell viability percentage (Viability%) using following formula:

CPD: the fluorescence signal value of compound well

ZPE (Zero percent effect): invalid effect control fluorescence signal value

Using following formula by original data processing be suppression percentage (Inhibition%):

CPD: the fluorescence signal value of compound well

HPE (Hundred percent effect): 100% useful effect compares fluorescence signal value

ZPE (Zero percent effect): invalid effect control fluorescence signal value

Suppression percentage importing GraphPad Prism is further processed and obtains homologous thread and EC₅₀Value.Data are shown in Table 2。

Table 2

Compound number	EC₅₀(nM)	Compound number	EC₅₀(nM)
				Positive control	80	Embodiment 1	113.6
Embodiment 2	32	Embodiment 3	60.46
				Embodiment 4	75.69	Embodiment 5	539.2
Embodiment 6	363.1	Embodiment 7	31
				Embodiment 8	316	Embodiment 9	>3330
Embodiment 10- I	291.1	Embodiment 10- II	1425

Embodiment 11	1475	Embodiment 12	638.4
				Embodiment 13	165.3	Embodiment 14	622.1
Embodiment 15	62.53
				Embodiment 16- I	42.52	Embodiment 16- II	260.1
Embodiment 17	192.9	Embodiment 18	577.1
				Embodiment 19	185.5	Embodiment 20	45
Embodiment 21	63	Embodiment 24	33
				Embodiment 25	37	Embodiment 26	68
Embodiment 27	150	Embodiment 28	58
				Embodiment 29	36	Embodiment 32	46
Embodiment 33	68

2 the compound of the present invention of experimental example detects the antiviral activity of HCV infection model (HCVcc) 2a

1 experimental material

1.1 compound

The compound of the present invention and control compound prepared by above embodiments 2,7 and 25 is configured to 10mM mother with DMSO After liquid, 10 μM are diluted to the DMEM complete culture solution containing 0.5%DMSO, then successively 3 times of dilutions, totally 10 concentration.

1.2 cell

Huh7.5.1 cell is provided by the bright Kant of medicine (Shanghai) new drug development Co., Ltd.

1.3 viral

HCVcc reporter virus has transfected the HCV overall length mutant strain of luciferase and GFP, can produce and JFH-1 Wild type has the virus of identical infection ability, is provided by the bright Kant of medicine (Shanghai) new drug development Co., Ltd.

1.4 reagent

DMEM cell culture fluid (DMEM medium) is purchased from U.S. Invitrogen company；

Fetal calf serum (Fetal bovine serum, FBS) is purchased from Sigma Co., USA；

L-Glutamine (L (+)-Glutamine) is purchased from U.S. Invitrogen company；

Pen .- Strep (Pen-Strep) is purchased from U.S. Invitrogen company；

Phosphate buffer (Phosphate buffered saline, PBS) is purchased from U.S. Hyclone company；

Pancreatin (Trypsin) is purchased from U.S. Invitrogen company；

Dimethyl sulfoxide (Dimethyl sulfoxide, DMSO) is purchased from Sigma Co., USA；

Cell pyrolysis liquid (lysis buffer) is purchased from U.S. Promega company；

Renillaluciferase detection reagent is purchased from U.S. Promega company；

Alamar Blue detection reagent is purchased from U.S. Invitrogen company.

1.4 instrument

EnVision multi-function microplate reader is purchased from U.S. Perkin-Elmer company.

2 experimental methods

1) Huh7.5.1 cell prepares: collecting the Huh7.5.1 cell of logarithmic phase, after being resuspended with DMEM complete culture solution, connects Kind is (7 × 10 in 96 orifice plates³A cells/well), 37 DEG C are placed in, 5%CO₂Overnight incubation in incubator；

2) after HCVcc reporter virus is resuspended with DMEM complete culture solution, 100 μ l viral supernatants (MOI virus infection: are added =0.2) into above-mentioned 96 orifice plate；

3) compound prepare: HCVcc reporter virus infection Huh7.5.1 cell in be added embodiment 2, embodiment 7, The compound of embodiment 25 and above-mentioned control compound, each each concentration of compound set duplicate hole；Invalid work is set up simultaneously With control group (Zero percent effect, ZPE) and 100% useful effect control group (Hundred percent Effect, HPE): ZPE group replaces compound with the complete culture solution containing 0.5%DMSO, and HPE group hole is the thin of virus-free infection Born of the same parents；

4) cell culture: 96 orifice plates are placed in 37 DEG C, 5%CO₂72hr is cultivated in incubator；

5) HCV-Ab IgG virus activity detects: after culture, discarding every hole supernatant, 20 μ l cell crackings are added into every hole Liquid and luciferase detection reagent read relative luminous intensity (RLU) with EnVision multi-function microplate reader, and initial data is used It is calculated in compound anti-HCV activity, calculation formula are as follows:

Inhibition%=(RLU_ZPE-RLU_CPD)/(RLU_ZPE-RLU_HPE)×100

Wherein RLU_CPDFor the fluorescence signal value for testing compound well, RLU_ZPEFor the fluorescence signal for acting on control wells in vain Value, RLU_HPEFor the fluorescence signal value of 100% useful effect control wells.

6) cell viability detects: planting the Huh7.5.1 cell into identical quantity in 96 orifice plates, and above-mentioned concentration ladder is added The compound of the present invention of degree handles 72hr；DMSO vehicle control group is set up simultaneously.After culture, 10% is added into every hole Alamar Blue detection reagent, 37 DEG C, 5%CO₂After incubator culture 2hr, read with EnVision multi-function microplate reader opposite Fluorescence intensity (RFU) is calculated using the data obtained for Compound Cytotoxicity, calculation formula are as follows:

Viability%=RFU_CPD/RFU_DMSO×100

7) data processing: Inhibition%, Viability% are directed respectively into GraphPad Prism software and counted According to processing, obtain compound to the medium effective concentration EC of HCVcc₅₀With half cytotoxic concentration CC₅₀, experimental result is shown in Table 3。

Table 3

Compound number	EC₅₀(nM)	CC₅₀(nM)	Compound number	EC₅₀(nM)	CC₅₀(nM)
						Positive control	12.18	>10000	Embodiment 2	2.074	>10000

Embodiment 7

1.604

>10000

Embodiment 25

0.615

>10000

From table 3 it can be seen that the compound of the embodiment of the present invention 2,7 and 25 has for HCV cell in vitro infection model Excellent antiviral activity, at the same it is small to cytotoxicity.

In addition, experiment, which is also shown that, uses HCV cell in vitro infection model, other chemical combination of the embodiment of the present invention preparation Object for example embodiment 1,5,6,8,9,10-I, 10-II, 11,12,13,14,15,16-I, 16-II, 17,18,19,20,21,22, 23,24,26,27,30,31,32,33,34,35 etc. compound has low medium effective concentration to HCVcc GT2a virus EC₅₀And high half cytotoxic concentration CC₅₀, show good inhibitory activity and small cytotoxicity.

The experimental results showed that, the compound of the present invention has the efficient ability for inhibiting HCV virus, with positive control above Medicine is compared, EC₅₀With suitable or superior effect, the prospect having had for treatment HCV infection.

Although being described in detail above to the present invention, however it is understood by skilled practitioners that without departing from this hair The present invention can be carry out various modifications and be changed under the premise of bright spirit and scope.Interest field of the invention is not limited to Detailed description made by above, and claims should be belonged to.

Claims

1. tricyclic fused heterocycle nucleoside phosphoramidate compound or its pharmaceutically acceptable salt, as shown in logical formula (I),

P* indicates chiral phosphorus atoms,

Wherein,

(1)R₁Selected from H and C_1-6Alkyl, the alkyl is optionally by one or more C_1-6Alkyl, C_1-6Alkoxy, C_1-6Alkylamino, halogen Element, hydroxyl, amino, nitro, cyano, C_1-6Alkyl acyl, aminoacyl, C_1-6Alkylamino acyl group, sulfydryl replace；

(2)R₂Selected from H, C_1-6Alkyl, aminoacyl, benzyl, the alkyl is optionally by one or more C_1-6Alkyl, C_1-6Alcoxyl Base, halogen, hydroxyl, amino, list C_1-6Alkylamino, double C_1-6Alkylamino replaces；

(3)R₃For there is no, or be selected from C_1-6Alkyl, C_1-6Alkoxy, fluorine, chlorine, bromine, halogenated C_1-6Alkyl, halogenated C_1-6Alkoxy, Hydroxyl, nitro, amino and-CN；

(4) D is selected from-O- ,-S- ,-SO- ,-SO₂-、-CO-、-N(R_b)-and-C (R_c1R_c2)-, the R_bTo be not present, hydrogen or C_1-6Alkyl, the R_c1、R_c2Respectively hydrogen, halogen, C_1-6Alkyl, C_2-6Alkenyl or halogenated C_2-6Alkenyl, or work as R_c1、R_c2It is When methyl, C can be with R_c1、R_c2Constitute C₃Naphthenic base；

(5)D¹Selected from-CO- and-(CH)_n, the n is 0；

(6)G、G¹Respectively-N- or-CH-；

(7)For singly-bound or double bond, two of themIt is singly-bound or oneFor singly-bound, and another For double bond；With

(8) ring T is selected from phenyl ring and C_3-8Cycloalkane, the phenyl ring or C_3-8Cycloalkane can be by R₄Replace, wherein R₄For C_1-6Alkane Base, C_2-6Alkenyl, C_2-6Alkynyl, C_1-6Alkoxy, halogen, halogenated C_1-6Alkyl, halogenated C_1-6Alkoxy, hydroxyl, nitro, amino, list C_1-6Alkylamino, double C_1-6Alkylamino, C_1-6Alkyl acyl, aminoacyl, C_1-6Alkylamino acyl group or-CN.

2. compound according to claim 1 or its pharmaceutically acceptable salt, wherein the R_bTo be not present, hydrogen, methyl, Ethyl, propyl, isopropyl, cyclopropyl, the R_c1、R_c2Respectively hydrogen, fluorine, methyl, ethyl, C_2-4Alkenyl or halogenated C_2-4Alkene Base, or work as R_c1、R_c2When being methyl, C can be with R_c1、R_c2Constitute cyclopropyl.

3. compound according to claim 1 or its pharmaceutically acceptable salt, middle ring T is selected from phenyl ring, pentamethylene, hexamethylene And cycloheptane.

4. the compound of -3 any one or its pharmaceutically acceptable salt according to claim 1, wherein D¹For there is no, then D, G、G¹Phenyl ring in connection constitutes five-membered ring fused benzene, as shown in logical formula (II),

5. a kind of compound or its pharmaceutically acceptable salt, the compound is the compound of structure in detail below:

6. pharmaceutical composition, it includes the compound of any one of claim 1-5 or its pharmaceutically acceptable salt and pharmacy Acceptable carrier.

7. the medicine group of the compound of -5 any one or its pharmaceutically acceptable salt or claim 6 according to claim 1 Object is closed, is used to treat flaviviridae infections.

8. the medicine group of the compound of -5 any one or its pharmaceutically acceptable salt or claim 6 according to claim 1 Object is closed, is used to treat infection with hepatitis C virus.

9. the pharmaceutical composition of the compound of any one of claim 1-5 or its pharmaceutically acceptable salt or claim 6 Application in the drug that preparation is used to treat and/or prevention of flavivirus coe virus infects.

10. application according to claim 9, wherein the flaviviridae is Hepatitis C Virus.